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Bio-Rad microseal
Microseal, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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microseal - by Bioz Stars, 2019-12
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Related Articles

Clone Assay:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Standard curves for absolute quantification constructed from Escherichia clones were prepared by diluting purified linearized plasmids containing our target of interest from 102 -109 gene copies. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Centrifugation:

Article Title: Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa
Article Snippet: First, 400 μL of FTA purification reagent was added to each well. .. The plate of samples was then covered with Microseal ‘F’ foil seals (Bio-Rad, Hercules, California, USA) to reduce contamination, vortexed, and incubated at room temperature for 4 min before centrifugation at 6200 rpm for 2 min at 4°C. .. The supernatant was removed and the plates were then centrifuged a second time before the foil was removed.

Amplification:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: Harvested amplified cDNAs from a 5 to 10 um C1 Fluidigm chip were transferred to a 96-well PCR plate (Denville, ThermoGrid PCR Plates 96-well standard, Cat# C18096-10). .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. Cycling conditions included an initial 2 minute denaturation at 98°C, followed by 40 cycles of 98°C for 2 s and annealing at 55°C for 2 s. A melt curve was created after the final cycle by increasing the temperature from 70–90°C in 0.2°C intervals and holding at each temperature for 10 s. qPCR reactions were performed in a CFX-96 Real-Time System (Bio-Rad, Hercules, CA).

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: The cDNA synthesis reaction proceeded at 50°C for 60 min. cDNA synthesis for each sample included a reaction lacking reverse transcriptase to test DNA contamination. qPCR consisted of 1X iTaq Universal SYBR Green Supermix (BioRad), 300 nM for both forward and reverse primers (final concentration) (IDT), and 100 ng cDNA template. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles. .. Data were analyzed by CFX Manager (BioRad) with FAM reporter and ROX as the reference dye.

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad). .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Positive Control:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: A positive control of seed DNA spiked with S. homoeocarpa genomic DNA and a no-template negative control were included in all runs and PCR was performed in a MasterCycle Pro S (Eppendorf, Hamburg, Germany). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Synthesized:

Article Title: Human Milk Oligosaccharide 2′-Fucosyllactose Improves Innate and Adaptive Immunity in an Influenza-Specific Murine Vaccination Model
Article Snippet: Before mRNA isolation, tissues were homogenized by using Precellys® CKMix Tissue Homogenizing Kit (Bertin, KT039611009.2) with the Precellys 24 homogenizer (Bertin, EQ 03119-200-RD000.0). mRNA was extracted using a NucleoSpin® RNA Plus kit (Macherey-Nagel, 740984.250) in combination with the rDNAse set (Macherey-Nagel, #740963) to remove contaminating DNA. cDNA was synthesized using an iScript™ advanced kit (Bio-Rad, Basel, Switzerland) according to the manufacturer’s protocol in the PTC-100 Programmable Thermal Controller (MJ research, PTC-100). cDNA and mRNA samples were stored at −80°C. .. The plate was covered with a Microseal “B” film (Bio-Rad, MSB1001) and then measured at the appropriate temperature for 40 cycles in the CFX96 Real-Time System C1000 Thermal Cycler (Bio-Rad, 1855195).

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: RT-qPCR cDNA was synthesized by incubating 5 μg of RNA, 250 ng of random primers, and 10 mM dNTP Mix (final concentration) at 65°C for 5 min followed by incubation on ice. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles.

Quantitative RT-PCR:

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: Eight-microlitre RT-qPCR mix containing 5.35 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer I (Applied Biosystems, #4379876), 1 μl 25 mM MgCl2 (Roche, #12032953001), 0.1 μl Rox Reference Dye (Invitrogen, #12223-012), 0.2 μl Assay On Demand (Applied Biosystems), 0.1 μl Multiscribe (50 U/μl) (Applied Biosystems, #4319983) and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. After sealing (Microseal ‘B’ film, Bio-Rad, #MSB1001), the plate was mixed and centrifuged for 3 min at 1450g .

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: The whole body section was placed within the matrix on the plate (tissue facing the well) and firmly sealed with a Microseal ‘B’ film (Bio-Rad, #MSB1001) with the help of a rubber brayer (Speedball). .. Lysates were transferred to a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) and stored at −80°C.

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: Paragraph title: Real-Time Quantitative PCR (RT-qPCR) ... Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles.

SYBR Green Assay:

Article Title: Human Milk Oligosaccharide 2′-Fucosyllactose Improves Innate and Adaptive Immunity in an Influenza-Specific Murine Vaccination Model
Article Snippet: Quantitative analysis was performed on a CFX96 Real-Time C1000 Thermal Cycler detection system with the use of an IQ™ SYBR® Green Supermix according to manufacturer’s protocol (both from Bio-Rad). .. The plate was covered with a Microseal “B” film (Bio-Rad, MSB1001) and then measured at the appropriate temperature for 40 cycles in the CFX96 Real-Time System C1000 Thermal Cycler (Bio-Rad, 1855195).

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: The cDNA synthesis reaction proceeded at 50°C for 60 min. cDNA synthesis for each sample included a reaction lacking reverse transcriptase to test DNA contamination. qPCR consisted of 1X iTaq Universal SYBR Green Supermix (BioRad), 300 nM for both forward and reverse primers (final concentration) (IDT), and 100 ng cDNA template. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles.

Article Title: Effect of IAPP on the proteome of cultured Rin-5F cells
Article Snippet: The PCR was performed in 10 μl of reaction volume with 5 μl of qPCR MasterMix Plus for SYBR® Green, 4 μl of 10x diluted cDNA and 0.1 μl of each forward and reverse primer at the stock concentration of 20 μM. .. The plates were sealed with Microseal ‘B’ film (Bio-Rad, UK) and centrifuged at 800 g for 1 min.

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Each 20-μL reaction aliquoted into the well of a Hard Shell PCR plate (Bio-Rad) contained 1X SsoAdvanced universal SYBR Green supermix (Bio-Rad), 0.5 μmol/L of final concentration of forward and reverse primers, and 2 μL of template DNA. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Incubation:

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged for 3 min at 1450g and incubated for 30 min at 33°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems). .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed and centrifuged for 3 min at 1450g .

Article Title: Low-input and multiplexed microfluidic assay reveals epigenomic variation across cerebellum and prefrontal cortex
Article Snippet: The microfluidic channel (with freshly oxidized glass surface) was filled with 0.1% PLL (P2636, Sigma-Aldrich) and sealed with Microseal ’B’ adhesive seals (MSB1001, Bio-Rad). .. Channel filling or washing was conducted by loading solution into a channel reservoir (or a 200-μl pipette tip plugged into the reservoir in the case of liquid volumes > 15 μl) on one end followed by aspiration using a pipettor on the other end (fig. S1B).

Article Title: Capture and characterization of influenza A virus from primary samples using glycan bead arrays
Article Snippet: IAV (50 µl) was added to the beads and the plate was sealed with microseal ‘F’ foil (Biorad). .. The plate was vortex-mixed and incubated at 4°C for 2h on a rocking shaker.

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: The cDNA synthesis reaction proceeded at 50°C for 60 min. cDNA synthesis for each sample included a reaction lacking reverse transcriptase to test DNA contamination. qPCR consisted of 1X iTaq Universal SYBR Green Supermix (BioRad), 300 nM for both forward and reverse primers (final concentration) (IDT), and 100 ng cDNA template. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles. .. Data were analyzed by CFX Manager (BioRad) with FAM reporter and ROX as the reference dye.

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl Chemical Ligation-mix containing 5.8 μl DEPC-water, 1 μl 10× PCR Buffer (Roche #14882500), 1 μl Poly(A) (1 μg/μl diluted in RNAse-free water/GE Healthcare, #27-4110-01), 0.1 μl 10 μM PS-Ligator (5′-TTAAACCATAGCAGCACG-PS-3′) and 0.1 μl 10 μM BPS-Ligator (5′-BPS-TAAATATTGGCGAACCAGT-3′) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged for 3 min at 1450g and incubated for 30 min at 33°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems). .. On completion of the chemical ligation reaction, 8 μl PCR-mix containing 6.375 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer (Roche #14882500), 0.15 μl 10 μM forward primer, 0.15 μl 10 μM reverse primer, 0.075 μl 50 μM Anti-primer and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to each well of a new 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2 μl of the Chemical Ligation-mix.

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl RT-mix containing 5.65 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer I (Applied Biosystems, #4379876), 1 μl 25 mM MgCl2 (Roche, #12032953001), 0.1 μl 1 μM RT-primer and 0.1 μl Multiscribe (50 U/μl) (Applied Biosystems, #4319983) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. The plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged and incubated for 10 min at 25°C followed by 5 min at 95°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems). .. Before unsealing, the plate was centrifuged for 3 min at 1450g , and 5 μl/well PCR mix was added.

Article Title: Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa
Article Snippet: First, 400 μL of FTA purification reagent was added to each well. .. The plate of samples was then covered with Microseal ‘F’ foil seals (Bio-Rad, Hercules, California, USA) to reduce contamination, vortexed, and incubated at room temperature for 4 min before centrifugation at 6200 rpm for 2 min at 4°C. .. The supernatant was removed and the plates were then centrifuged a second time before the foil was removed.

Activity Assay:

Article Title: Capture and characterization of influenza A virus from primary samples using glycan bead arrays
Article Snippet: IAV (50 µl) was added to the beads and the plate was sealed with microseal ‘F’ foil (Biorad). .. The plate was vortex-mixed and incubated at 4°C for 2h on a rocking shaker.

Expressing:

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles.

Transformation Assay:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. The plasmid used for generation of standard curves and copy number estimation was constructed by cloning S. homoeocarpa EF1α sequence amplified by our primers into a pGEM-T Easy plasmid (Promega, Madison, WI) according to the manufacturer's instructions.

Blocking Assay:

Article Title: Low-input and multiplexed microfluidic assay reveals epigenomic variation across cerebellum and prefrontal cortex
Article Snippet: The microfluidic channel (with freshly oxidized glass surface) was filled with 0.1% PLL (P2636, Sigma-Aldrich) and sealed with Microseal ’B’ adhesive seals (MSB1001, Bio-Rad). .. The microfluidic channel (with freshly oxidized glass surface) was filled with 0.1% PLL (P2636, Sigma-Aldrich) and sealed with Microseal ’B’ adhesive seals (MSB1001, Bio-Rad).

Article Title: Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa
Article Snippet: A 96-well, Square V-Bottom 2-mL Assay Block (Costar, Corning Inc., Corning, New York, USA) was used with eight punched disks placed in each well. .. The plate of samples was then covered with Microseal ‘F’ foil seals (Bio-Rad, Hercules, California, USA) to reduce contamination, vortexed, and incubated at room temperature for 4 min before centrifugation at 6200 rpm for 2 min at 4°C.

Flow Cytometry:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001). .. The concentration of amplified cDNA was measured by using the Quant-iT™ PicoGreen® dsDNA Assay Kit (Invitrogen Cat# P11496).

Ligation:

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: On completion of the chemical ligation reaction, 8 μl PCR-mix containing 6.375 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer (Roche #14882500), 0.15 μl 10 μM forward primer, 0.15 μl 10 μM reverse primer, 0.075 μl 50 μM Anti-primer and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to each well of a new 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2 μl of the Chemical Ligation-mix. .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed and centrifuged for 3 min at 1450g .

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl Chemical Ligation-mix containing 5.8 μl DEPC-water, 1 μl 10× PCR Buffer (Roche #14882500), 1 μl Poly(A) (1 μg/μl diluted in RNAse-free water/GE Healthcare, #27-4110-01), 0.1 μl 10 μM PS-Ligator (5′-TTAAACCATAGCAGCACG-PS-3′) and 0.1 μl 10 μM BPS-Ligator (5′-BPS-TAAATATTGGCGAACCAGT-3′) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged for 3 min at 1450g and incubated for 30 min at 33°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems).

Generated:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001). .. The concentration of amplified cDNA was measured by using the Quant-iT™ PicoGreen® dsDNA Assay Kit (Invitrogen Cat# P11496).

Inhibition:

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Endogenous positive controls (pure extracted DNA of the targeted species) were run to assess PCR inhibition and determination of specific melt curve temperatures for data analysis. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

DNA Sequencing:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Positive bands were excised from the gel with a sterile razor blade and cleaned up with the Wizard SV Gel and PCR Clean-Up System (Promega, Madison, WI) then sequenced at the University of Wisconsin-Madison DNA Sequencing facility (Madison, WI). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Sequencing:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001). .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: NCBI BLASTn was used to confirm sequences were specific to the S. homoeocarpa EF1- α gene sequence. .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Binding Assay:

Article Title: Capture and characterization of influenza A virus from primary samples using glycan bead arrays
Article Snippet: Paragraph title: Virus binding to 3D mucin mimetic array ... IAV (50 µl) was added to the beads and the plate was sealed with microseal ‘F’ foil (Biorad).

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad). .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Imaging:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Gels were visualized with UV light in a Universal Hood II Gel Doc System (BioRad, Hercules, CA) and analyzed with Quantity One imaging software (BioRad, Hercules, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

RNA Sequencing Assay:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: Paragraph title: Single cell RNA-seq using the fluidigm C1 system ... The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

Fluorescence:

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: Paragraph title: Isolation of single human B cells by Fluorescence Activated Cell Sorting ... Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

Magnetic Beads:

Article Title: Capture and characterization of influenza A virus from primary samples using glycan bead arrays
Article Snippet: All washes were done on a DynaMag-96 side skirted magnetic beads separator plate (Life Technologies), the PCR plate was shifted back and forth from left to right 10 times to mix the beads. .. IAV (50 µl) was added to the beads and the plate was sealed with microseal ‘F’ foil (Biorad).

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: If necessary, for example due to low B cell counts in samples from patients with autoimmune disease, B cells were enriched using anti-CD19 magnetic beads (Miltenyi Biotech). .. Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

Isolation:

Article Title: Human Milk Oligosaccharide 2′-Fucosyllactose Improves Innate and Adaptive Immunity in an Influenza-Specific Murine Vaccination Model
Article Snippet: Before mRNA isolation, tissues were homogenized by using Precellys® CKMix Tissue Homogenizing Kit (Bertin, KT039611009.2) with the Precellys 24 homogenizer (Bertin, EQ 03119-200-RD000.0). mRNA was extracted using a NucleoSpin® RNA Plus kit (Macherey-Nagel, 740984.250) in combination with the rDNAse set (Macherey-Nagel, #740963) to remove contaminating DNA. cDNA was synthesized using an iScript™ advanced kit (Bio-Rad, Basel, Switzerland) according to the manufacturer’s protocol in the PTC-100 Programmable Thermal Controller (MJ research, PTC-100). cDNA and mRNA samples were stored at −80°C. .. The plate was covered with a Microseal “B” film (Bio-Rad, MSB1001) and then measured at the appropriate temperature for 40 cycles in the CFX96 Real-Time System C1000 Thermal Cycler (Bio-Rad, 1855195).

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: Paragraph title: Isolation of single human B cells by Fluorescence Activated Cell Sorting ... Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

Negative Control:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: A positive control of seed DNA spiked with S. homoeocarpa genomic DNA and a no-template negative control were included in all runs and PCR was performed in a MasterCycle Pro S (Eppendorf, Hamburg, Germany). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Microscopy:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: After cell counting we loaded cells at a concentration of 2.5 × 105 /ml on a C1 Fluidigm 5–10 um RNA-seq chip immediately, and the number of captured cells at each capture site was recorded under a microscope. .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

Purification:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001). .. The libraries were generated by using a Nextera XT DNA sample preparation kit (96 samples) (Illumina, Catalog# FC-131-1096) and Truseq Dual Index sequencing Primer (paired-end) kit (Illumina, Catalog# PE-121-1003), with 96 barcoded samples combined.

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Article Title: Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa
Article Snippet: First, 400 μL of FTA purification reagent was added to each well. .. The plate of samples was then covered with Microseal ‘F’ foil seals (Bio-Rad, Hercules, California, USA) to reduce contamination, vortexed, and incubated at room temperature for 4 min before centrifugation at 6200 rpm for 2 min at 4°C.

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: Purified mononuclear cells were stained on ice with anti-human antibodies (Becton Dickinson) directly coupled to fluorescein-isothiocyanate (FITC), phycoerythrin (PE), allophycocyanin (APC), or biotin to distinguish among individual B cell subpopulations ( ). .. Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Standard curves for absolute quantification constructed from Escherichia clones were prepared by diluting purified linearized plasmids containing our target of interest from 102 -109 gene copies. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Polymerase Chain Reaction:

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: Eight-microlitre RT-qPCR mix containing 5.35 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer I (Applied Biosystems, #4379876), 1 μl 25 mM MgCl2 (Roche, #12032953001), 0.1 μl Rox Reference Dye (Invitrogen, #12223-012), 0.2 μl Assay On Demand (Applied Biosystems), 0.1 μl Multiscribe (50 U/μl) (Applied Biosystems, #4319983) and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. After sealing (Microseal ‘B’ film, Bio-Rad, #MSB1001), the plate was mixed and centrifuged for 3 min at 1450g .

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: On completion of the chemical ligation reaction, 8 μl PCR-mix containing 6.375 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer (Roche #14882500), 0.15 μl 10 μM forward primer, 0.15 μl 10 μM reverse primer, 0.075 μl 50 μM Anti-primer and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to each well of a new 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2 μl of the Chemical Ligation-mix. .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed and centrifuged for 3 min at 1450g .

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: The whole body section was placed within the matrix on the plate (tissue facing the well) and firmly sealed with a Microseal ‘B’ film (Bio-Rad, #MSB1001) with the help of a rubber brayer (Speedball). .. The whole body section was placed within the matrix on the plate (tissue facing the well) and firmly sealed with a Microseal ‘B’ film (Bio-Rad, #MSB1001) with the help of a rubber brayer (Speedball).

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: Harvested amplified cDNAs from a 5 to 10 um C1 Fluidigm chip were transferred to a 96-well PCR plate (Denville, ThermoGrid PCR Plates 96-well standard, Cat# C18096-10). .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001). .. The concentration of amplified cDNA was measured by using the Quant-iT™ PicoGreen® dsDNA Assay Kit (Invitrogen Cat# P11496).

Article Title: Capture and characterization of influenza A virus from primary samples using glycan bead arrays
Article Snippet: All washes were done on a DynaMag-96 side skirted magnetic beads separator plate (Life Technologies), the PCR plate was shifted back and forth from left to right 10 times to mix the beads. .. IAV (50 µl) was added to the beads and the plate was sealed with microseal ‘F’ foil (Biorad).

Article Title: Human Milk Oligosaccharide 2′-Fucosyllactose Improves Innate and Adaptive Immunity in an Influenza-Specific Murine Vaccination Model
Article Snippet: 300 nM forward and reverse primers were used in 10 µL reactions (8.8 µL mix + 1.2 µL cDNA), which were measured in a 96 well Hard-Shell PCR plate with thin walls (Bio-Rad, HSP9601). .. The plate was covered with a Microseal “B” film (Bio-Rad, MSB1001) and then measured at the appropriate temperature for 40 cycles in the CFX96 Real-Time System C1000 Thermal Cycler (Bio-Rad, 1855195).

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: NCBI BLASTn was used to confirm sequences were specific to the S. homoeocarpa EF1- α gene sequence. .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. A standard curve with concentrations of EF1α plasmid DNA ranging from 106 to one copy/µL was included on each plate and all DNA samples were assayed in triplicate.

Article Title: Effect of IAPP on the proteome of cultured Rin-5F cells
Article Snippet: The PCR was performed in 10 μl of reaction volume with 5 μl of qPCR MasterMix Plus for SYBR® Green, 4 μl of 10x diluted cDNA and 0.1 μl of each forward and reverse primer at the stock concentration of 20 μM. .. The plates were sealed with Microseal ‘B’ film (Bio-Rad, UK) and centrifuged at 800 g for 1 min.

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl Chemical Ligation-mix containing 5.8 μl DEPC-water, 1 μl 10× PCR Buffer (Roche #14882500), 1 μl Poly(A) (1 μg/μl diluted in RNAse-free water/GE Healthcare, #27-4110-01), 0.1 μl 10 μM PS-Ligator (5′-TTAAACCATAGCAGCACG-PS-3′) and 0.1 μl 10 μM BPS-Ligator (5′-BPS-TAAATATTGGCGAACCAGT-3′) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged for 3 min at 1450g and incubated for 30 min at 33°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems).

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl RT-mix containing 5.65 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer I (Applied Biosystems, #4379876), 1 μl 25 mM MgCl2 (Roche, #12032953001), 0.1 μl 1 μM RT-primer and 0.1 μl Multiscribe (50 U/μl) (Applied Biosystems, #4319983) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. The plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged and incubated for 10 min at 25°C followed by 5 min at 95°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems).

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: Single cells were sorted on a FACSVantage (Becton Dickinson) excluding cell duplets into 96-well PCR plates (Eppendorf) containing 4 μl/well of ice-cold 0.5× phosphate-buffered saline (PBS) containing 10 mM DTT, 8 U RNAsin (Promega), 0.4 U 5′-3′ Prime RNAse Inhibitor (Eppendorf). .. Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Each 20-μL reaction aliquoted into the well of a Hard Shell PCR plate (Bio-Rad) contained 1X SsoAdvanced universal SYBR Green supermix (Bio-Rad), 0.5 μmol/L of final concentration of forward and reverse primers, and 2 μL of template DNA. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Construct:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. Cycling conditions included an initial 2 minute denaturation at 98°C, followed by 40 cycles of 98°C for 2 s and annealing at 55°C for 2 s. A melt curve was created after the final cycle by increasing the temperature from 70–90°C in 0.2°C intervals and holding at each temperature for 10 s. qPCR reactions were performed in a CFX-96 Real-Time System (Bio-Rad, Hercules, CA).

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Standard curves for absolute quantification constructed from Escherichia clones were prepared by diluting purified linearized plasmids containing our target of interest from 102 -109 gene copies. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Staining:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Five µL of all primary and nested PCR products were mixed with 1 µL of 6× DNA loading dye and subject to electrophoresis in a 1% agarose gel stained with SYBR Safe DNA Gel Stain (Invitrogen, Carlsbad, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: Purified mononuclear cells were stained on ice with anti-human antibodies (Becton Dickinson) directly coupled to fluorescein-isothiocyanate (FITC), phycoerythrin (PE), allophycocyanin (APC), or biotin to distinguish among individual B cell subpopulations ( ). .. Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

Nested PCR:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Five µL of all primary and nested PCR products were mixed with 1 µL of 6× DNA loading dye and subject to electrophoresis in a 1% agarose gel stained with SYBR Safe DNA Gel Stain (Invitrogen, Carlsbad, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Sample Prep:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001). .. The concentration of amplified cDNA was measured by using the Quant-iT™ PicoGreen® dsDNA Assay Kit (Invitrogen Cat# P11496).

Mouse Assay:

Article Title: Human Milk Oligosaccharide 2′-Fucosyllactose Improves Innate and Adaptive Immunity in an Influenza-Specific Murine Vaccination Model
Article Snippet: qPCR Analysis Ileum samples from influenza-vaccinated mice sacrificed after DTH measurement were collected in RNAlater (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) and over night stored in the fridge, after which they were stored at −80°C until mRNA isolation. .. The plate was covered with a Microseal “B” film (Bio-Rad, MSB1001) and then measured at the appropriate temperature for 40 cycles in the CFX96 Real-Time System C1000 Thermal Cycler (Bio-Rad, 1855195).

Chromatin Immunoprecipitation:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: Harvested amplified cDNAs from a 5 to 10 um C1 Fluidigm chip were transferred to a 96-well PCR plate (Denville, ThermoGrid PCR Plates 96-well standard, Cat# C18096-10). .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

Plasmid Preparation:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: NCBI BLASTn was used to confirm sequences were specific to the S. homoeocarpa EF1- α gene sequence. .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. A standard curve with concentrations of EF1α plasmid DNA ranging from 106 to one copy/µL was included on each plate and all DNA samples were assayed in triplicate.

Software:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Gels were visualized with UV light in a Universal Hood II Gel Doc System (BioRad, Hercules, CA) and analyzed with Quantity One imaging software (BioRad, Hercules, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Real-time Polymerase Chain Reaction:

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged for 3 min at 1450g and incubated for 30 min at 33°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems). .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed and centrifuged for 3 min at 1450g .

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: Eight-microlitre qPCR mix containing 5.45 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer I (Applied Biosystems, #4379876), 1 μl 25 mM MgCl2 (Roche, #12032953001), 0.1 μl Rox Reference Dye (Invitrogen, #12223-012), 0.2 μl Assay On Demand (Applied Biosystems) and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. After sealing (Microseal ‘B’ film, Bio-Rad, #MSB1001), the plate was mixed and centrifuged for 3 min at 1450g .

Article Title: Human Milk Oligosaccharide 2′-Fucosyllactose Improves Innate and Adaptive Immunity in an Influenza-Specific Murine Vaccination Model
Article Snippet: Paragraph title: qPCR Analysis ... The plate was covered with a Microseal “B” film (Bio-Rad, MSB1001) and then measured at the appropriate temperature for 40 cycles in the CFX96 Real-Time System C1000 Thermal Cycler (Bio-Rad, 1855195).

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: NCBI BLASTn was used to confirm sequences were specific to the S. homoeocarpa EF1- α gene sequence. .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. A standard curve with concentrations of EF1α plasmid DNA ranging from 106 to one copy/µL was included on each plate and all DNA samples were assayed in triplicate.

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: The cDNA synthesis reaction proceeded at 50°C for 60 min. cDNA synthesis for each sample included a reaction lacking reverse transcriptase to test DNA contamination. qPCR consisted of 1X iTaq Universal SYBR Green Supermix (BioRad), 300 nM for both forward and reverse primers (final concentration) (IDT), and 100 ng cDNA template. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles. .. Data were analyzed by CFX Manager (BioRad) with FAM reporter and ROX as the reference dye.

Article Title: Effect of IAPP on the proteome of cultured Rin-5F cells
Article Snippet: Paragraph title: Quantitative real time PCR ... The plates were sealed with Microseal ‘B’ film (Bio-Rad, UK) and centrifuged at 800 g for 1 min.

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl Chemical Ligation-mix containing 5.8 μl DEPC-water, 1 μl 10× PCR Buffer (Roche #14882500), 1 μl Poly(A) (1 μg/μl diluted in RNAse-free water/GE Healthcare, #27-4110-01), 0.1 μl 10 μM PS-Ligator (5′-TTAAACCATAGCAGCACG-PS-3′) and 0.1 μl 10 μM BPS-Ligator (5′-BPS-TAAATATTGGCGAACCAGT-3′) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. Subsequently, the plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged for 3 min at 1450g and incubated for 30 min at 33°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems). .. On completion of the chemical ligation reaction, 8 μl PCR-mix containing 6.375 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer (Roche #14882500), 0.15 μl 10 μM forward primer, 0.15 μl 10 μM reverse primer, 0.075 μl 50 μM Anti-primer and 0.1 μl Hot Start Taq Polymerase (5 U/μl) (Roche, #12032953001) was added to each well of a new 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2 μl of the Chemical Ligation-mix.

Article Title: Whole-body scanning PCR; a highly sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides
Article Snippet: In the first step, 8 μl RT-mix containing 5.65 μl DEPC-water, 0.15 μl 100 mM dNTPs (Applied Biosystems, #362271), 1 μl 10× PCR Buffer I (Applied Biosystems, #4379876), 1 μl 25 mM MgCl2 (Roche, #12032953001), 0.1 μl 1 μM RT-primer and 0.1 μl Multiscribe (50 U/μl) (Applied Biosystems, #4319983) was added to each well of a 384-well Hard-Shell PCR Plate (Bio-Rad, #HSP3901) followed by the addition of 2-μl sample. .. The plate was sealed (Microseal ‘B’ film, Bio-Rad, #MSB1001), mixed, centrifuged and incubated for 10 min at 25°C followed by 5 min at 95°C in a 7900HT Fast Real-Time PCR System (Applied Biosystems). .. Before unsealing, the plate was centrifuged for 3 min at 1450g , and 5 μl/well PCR mix was added.

Article Title: The stability of an mRNA is influenced by its concentration: a potential physical mechanism to regulate gene expression
Article Snippet: Synthesis of cDNA was performed on 10 μg of total RNAs using SuperScript II reverse transcriptase (Life Technologies) as previously described ( ). .. The cDNAs were 1:10 serially diluted and the qPCR was performed in a CFX96 Real Time PCR Detection System (Bio-Rad), using 96-well plate (Bio-Rad) sealed with Microseal ‘B’ seals (Bio-Rad) as previously described ( ). .. Diluted cDNA from cultures in steady state (without transcription arrest with rifampicin) were used to identify the dilution that would lead to a cycle threshold (Ct) of between 15 and 25.

RNA Extraction:

Article Title: The stability of an mRNA is influenced by its concentration: a potential physical mechanism to regulate gene expression
Article Snippet: Paragraph title: RNA extraction, quality control and cDNA synthesis ... The cDNAs were 1:10 serially diluted and the qPCR was performed in a CFX96 Real Time PCR Detection System (Bio-Rad), using 96-well plate (Bio-Rad) sealed with Microseal ‘B’ seals (Bio-Rad) as previously described ( ).

Agarose Gel Electrophoresis:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Five µL of all primary and nested PCR products were mixed with 1 µL of 6× DNA loading dye and subject to electrophoresis in a 1% agarose gel stained with SYBR Safe DNA Gel Stain (Invitrogen, Carlsbad, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Electrophoresis:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Five µL of all primary and nested PCR products were mixed with 1 µL of 6× DNA loading dye and subject to electrophoresis in a 1% agarose gel stained with SYBR Safe DNA Gel Stain (Invitrogen, Carlsbad, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

Spectrophotometry:

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: RNA concentrations were determined using a NanoDrop Spectrophotometer ND-1000. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles.

Article Title: The stability of an mRNA is influenced by its concentration: a potential physical mechanism to regulate gene expression
Article Snippet: RNAs were quantified using ND-1000 UV-visible light spectrophotometer (NanoDrop Technologies) and their integrity certified with Bioanalyzer 2100 with the RNA 6000 Nano LabChip kit (Agilent). .. The cDNAs were 1:10 serially diluted and the qPCR was performed in a CFX96 Real Time PCR Detection System (Bio-Rad), using 96-well plate (Bio-Rad) sealed with Microseal ‘B’ seals (Bio-Rad) as previously described ( ).

Concentration Assay:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: After cell counting we loaded cells at a concentration of 2.5 × 105 /ml on a C1 Fluidigm 5–10 um RNA-seq chip immediately, and the number of captured cells at each capture site was recorded under a microscope. .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

Article Title: A Dual Role for the Bacillus anthracis Master Virulence Regulator AtxA: Control of Sporulation and Anthrax Toxin Production
Article Snippet: The cDNA synthesis reaction proceeded at 50°C for 60 min. cDNA synthesis for each sample included a reaction lacking reverse transcriptase to test DNA contamination. qPCR consisted of 1X iTaq Universal SYBR Green Supermix (BioRad), 300 nM for both forward and reverse primers (final concentration) (IDT), and 100 ng cDNA template. .. Each qPCR plate contained a no-template control for each sample to ensure reagents were not contaminated. qPCR plates were covered with Microseal “C” Film (BioRad) and incubated in a CFX96 Real Time PCR Detection System (BioRad) using the following cycling conditions: 95°C, 2 min; followed by 40 cycles of 95°C, 15 s and 60°C, 30 s. Melt curve analysis (65°C–95°C at 0.5°C increments for 2–5 s/step) was performed at the conclusion of amplification cycles.

Article Title: Effect of IAPP on the proteome of cultured Rin-5F cells
Article Snippet: The PCR was performed in 10 μl of reaction volume with 5 μl of qPCR MasterMix Plus for SYBR® Green, 4 μl of 10x diluted cDNA and 0.1 μl of each forward and reverse primer at the stock concentration of 20 μM. .. The plates were sealed with Microseal ‘B’ film (Bio-Rad, UK) and centrifuged at 800 g for 1 min.

Article Title: Impact of contraceptive initiation on vaginal microbiota
Article Snippet: Each 20-μL reaction aliquoted into the well of a Hard Shell PCR plate (Bio-Rad) contained 1X SsoAdvanced universal SYBR Green supermix (Bio-Rad), 0.5 μmol/L of final concentration of forward and reverse primers, and 2 μL of template DNA. .. Plates were sealed with Microseal “B” adhesive optically clear seals (Bio-Rad).

Cell Counting:

Article Title: Single cell transcriptomics reveals unanticipated features of early hematopoietic precursors
Article Snippet: After cell counting we loaded cells at a concentration of 2.5 × 105 /ml on a C1 Fluidigm 5–10 um RNA-seq chip immediately, and the number of captured cells at each capture site was recorded under a microscope. .. The PCR plate was sealed with Microseal ‘B’ seal Seals (Bio-rad Cat# MSB1001).

DNA Purification:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL. .. The plasmid used for generation of standard curves and copy number estimation was constructed by cloning S. homoeocarpa EF1α sequence amplified by our primers into a pGEM-T Easy plasmid (Promega, Madison, WI) according to the manufacturer's instructions.

FACS:

Article Title: Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
Article Snippet: Paragraph title: Isolation of single human B cells by Fluorescence Activated Cell Sorting ... Plates were sealed with Microseal® ‘F’ Film (BioRad) and immediately frozen on dry ice before storage at −80°C.

other:

Article Title: Low-input and multiplexed microfluidic assay reveals epigenomic variation across cerebellum and prefrontal cortex
Article Snippet: PLL/oligo linker Our antibody coating procedure was similar to previous works with minor modifications ( , ).

Hood:

Article Title: Sclerotinia homoeocarpa Overwinters in Turfgrass and Is Present in Commercial Seed
Article Snippet: Gels were visualized with UV light in a Universal Hood II Gel Doc System (BioRad, Hercules, CA) and analyzed with Quantity One imaging software (BioRad, Hercules, CA). .. Quantitative PCR (qPCR) reactions were performed in hard-shell 96-well skirted PCR plates and sealed with Microseal ‘B’ adhesive seals (BioRad, Hercules, CA). qPCR reactions contained 1× SsoFast EvaGreen Supermix, 8 µL template (seed DNA or EF1α plasmid DNA standard), 0.2 µM EF1α_nest_F and EF1α_nest_R, and NFW for a final reaction volume of 20 µL.

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    Bio-Rad microwells
    Arrays of live-imaged embryos within the OMMAwell mold. (A) A single timepoint from a time-lapse of an array of nuclear-marked transgenic cricket embryos in <t>microwells.</t> The two leftmost columns show germ band stage embryos that are beginning the physical re-orientation within the egg called anatrepsis. The two rightmost columns show a later stage when embryos are fully immersed in the yolk below the extraembryonic membrane called the serosa. (B) Time series of cricket embryos starting at different ages. The ambient temperature is ∼24°C, so development is slower than that reported by Donoughe and Extavour (2016) . Scale bars: 500 µm.
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    82
    Bio-Rad high performance microbeads 27 plex assay
    Overview of serum chemokines, cytokines and growth factors of the atrial fibrillation patient compared to control groups. Serum biomarkers: CXCL8 (IL-8); CXCL10 (IP-10); CCL11 (Eotaxin); CCL3 (MIP-1α); CCL4 (MIP-1β); CCL2 (MCP-1); CCL-5 (RANTES); IL1-β, IL-6, TNF-α, IL-12; IFN-γ, IL-17; IL-1Ra (IL-1 receptor antagonist); IL-2; IL-4; IL-5; IL-7; IL-9; IL-10; IL-13; IL-15; FGF-basic; PDGF; VEGF; G-CSF and GM-CSF were measured by high performance microbeads 27-plex assay, as described in materials and methods, with the 95%CI represented by gray boxes. Black arrows represent an increase (up) or decrease (down) in the patient’s biomarkers level in comparison to control groups; a black square represents biomarkers levels at the 95%CI.  a  Biomarkers’ levels: case vs. gender matching non-infected heath controls; ( b ) Biomarkers’ levels: case vs. gender matching ZIKV-infected patients
    High Performance Microbeads 27 Plex Assay, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 82/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Bio-Rad microbead containing tubes
    A MR100-based assay can differentiate between prion-infected and normal brain homogenates without proteinase K digestion. a Schematic description of the RCA protocol to test brain homogenates without PK digestion. Brain tissues were freshly homogenized in <t>microbead-containing</t> tubes. Normal brain homogenates (NBH) or prion-infected brain homogenates (IBH) were incubated with MR100 for 1 h, at room temperature, leading to a precipitation of PrP isoforms. After a short centrifugation step, the pellet with MR100 (orange tube) concentrates PrP isoforms, whereas no pellet is detectable with DMSO. b Comparison of DMSO, P30 and MR100 precipitation capabilities using the RCA protocol. Fifty μL of 10 % 22 L infected brain homogenates were diluted in 300 μL of PBS/2 % sarcosyl and incubated using either 1.5 mM of P30 or MR100 or an equivalent volume of the solvent alone (DMSO), at room temperature for 1 h. Then, samples were centrifuged at 8000 g for 5 min. Supernatants (S) were collected and 30 μL of each supernatant was mixed with an equivalent volume of 2X loading buffer. Pellets (P) were resuspended in 30 μL PBS/2 % Sarcosyl, and mixed with an equal volume of 2X loading buffer. Thirty microliters of each sample were loaded on 12 % Bis-Tris gels (Criterion, Biorad) and immunoblotting was carried out with the SAF mix according to standard procedures [ 16 ]. The samples (S/P) were analyzed by western blotting using SAF mix anti-PrP antibodies. c Comparison of infected versus non-infected brain homogenates processed with the RCA protocol. Fifty microliters of 10 % freshly homogenized brain tissues from normal (NBH) or 22 L prion-infected (IBH) mice were processed according to the RCA protocol described in A and B. Thirty microliters of supernatant (S) or pellet (P) were loaded on 12 % Bis-Tris gels (Criterion, Biorad) and immunoblotting was carried out with the SAF mix as described above. Molecular masses (20–75 kDa) are indicated on the left side of the panels
    Microbead Containing Tubes, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 87/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad elisa microwell plates
    Biochemical properties of SVmab and rSVmab. A Binding activity of SVmab and rSVmab in <t>ELISA.</t> hVSD II (1 μg) was coated on the <t>microwell</t> plate in 0.05 mmol/L carconate-bicarbonate buffer, pH 9.6. Anti-FLAG antibody was used as a negative control. All ELISA measurements were performed in triplicate. Data are mean ± SEM. B Antibody binding in ELISA. SVmab (66 nmol/L), SVmab (first batch, 66 nmol/L), and CTmab (66 nmol/L) were used. Anti-FLAG antibody (266 nmol/L) was added as a positive control. All ELISA measurements were performed in triplicate. Data are mean ± SEM. C SDS-PAGE (left) and western blots (right) of SVmab (2 μg) and rSVmab (2 μg) under a non-reducing condition. The gel (left) was stained with Coomassie blue. D SDS-PAGE of SVmab (1 μg) and rSVmab (1 μg) under a reducing condition. The gel was stained with Coomassie blue.
    Elisa Microwell Plates, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Arrays of live-imaged embryos within the OMMAwell mold. (A) A single timepoint from a time-lapse of an array of nuclear-marked transgenic cricket embryos in microwells. The two leftmost columns show germ band stage embryos that are beginning the physical re-orientation within the egg called anatrepsis. The two rightmost columns show a later stage when embryos are fully immersed in the yolk below the extraembryonic membrane called the serosa. (B) Time series of cricket embryos starting at different ages. The ambient temperature is ∼24°C, so development is slower than that reported by Donoughe and Extavour (2016) . Scale bars: 500 µm.

    Journal: Biology Open

    Article Title: High-throughput live-imaging of embryos in microwell arrays using a modular specimen mounting system

    doi: 10.1242/bio.031260

    Figure Lengend Snippet: Arrays of live-imaged embryos within the OMMAwell mold. (A) A single timepoint from a time-lapse of an array of nuclear-marked transgenic cricket embryos in microwells. The two leftmost columns show germ band stage embryos that are beginning the physical re-orientation within the egg called anatrepsis. The two rightmost columns show a later stage when embryos are fully immersed in the yolk below the extraembryonic membrane called the serosa. (B) Time series of cricket embryos starting at different ages. The ambient temperature is ∼24°C, so development is slower than that reported by Donoughe and Extavour (2016) . Scale bars: 500 µm.

    Article Snippet: When making the microwells, agarose (Bio-Rad #1613101) was dissolved at 1.5% weight/volume (w/v) in distilled water (or 2% for firmer molds).

    Techniques: Transgenic Assay

    Designing microwells to hold cricket eggs. (A) Freshly laid cricket eggs were measured and their (B) lengths and (C) widths plotted ( n =98; scale bar: 500 µm). Based on the size distribution, we chose dimensions of 2930×570×650 µm (embryo length×width×height), which were values such that approximately 75% of eggs would fit into the troughs. In practice, because the wells are made of agarose, more than 95% of eggs fit into these wells. (D) Raised posts of those dimensions were formed by engraving an acrylic insert. 120 such posts were arranged in a grid. (E) Agarose microwells made using this insert, loaded with cricket eggs. (F,G) Example molds for an annelid worm and coqui frog, with microwell dimensions listed. Photos in F and G by Elaine Seaver (The Whitney Laboratory for Marine Bioscience) and Mara Laslo (Harvard University) respectively.

    Journal: Biology Open

    Article Title: High-throughput live-imaging of embryos in microwell arrays using a modular specimen mounting system

    doi: 10.1242/bio.031260

    Figure Lengend Snippet: Designing microwells to hold cricket eggs. (A) Freshly laid cricket eggs were measured and their (B) lengths and (C) widths plotted ( n =98; scale bar: 500 µm). Based on the size distribution, we chose dimensions of 2930×570×650 µm (embryo length×width×height), which were values such that approximately 75% of eggs would fit into the troughs. In practice, because the wells are made of agarose, more than 95% of eggs fit into these wells. (D) Raised posts of those dimensions were formed by engraving an acrylic insert. 120 such posts were arranged in a grid. (E) Agarose microwells made using this insert, loaded with cricket eggs. (F,G) Example molds for an annelid worm and coqui frog, with microwell dimensions listed. Photos in F and G by Elaine Seaver (The Whitney Laboratory for Marine Bioscience) and Mara Laslo (Harvard University) respectively.

    Article Snippet: When making the microwells, agarose (Bio-Rad #1613101) was dissolved at 1.5% weight/volume (w/v) in distilled water (or 2% for firmer molds).

    Techniques:

    OMMAwell configurations for bottom-loaded microwells. Configuration 3: Bottom loaded microwells with a reservoir of live-imaging medium. (1) The insert (green) and cylinder (purple) are placed into the sheath (orange). (2) Molten agarose is poured into the cylinder to the desired depth. (3) Once the agarose has set, the cylinder and the agarose block are removed from the sheath and insert. The cylinder is flipped over, and the exposed microwells are loaded with embryos, as described in Fig. 2 . (4) The cylinder and agarose block are lowered into the glass-bottom dish, and live-imaging medium is added in the cylinder. Right: Schematic of embryo in Configuration 3.

    Journal: Biology Open

    Article Title: High-throughput live-imaging of embryos in microwell arrays using a modular specimen mounting system

    doi: 10.1242/bio.031260

    Figure Lengend Snippet: OMMAwell configurations for bottom-loaded microwells. Configuration 3: Bottom loaded microwells with a reservoir of live-imaging medium. (1) The insert (green) and cylinder (purple) are placed into the sheath (orange). (2) Molten agarose is poured into the cylinder to the desired depth. (3) Once the agarose has set, the cylinder and the agarose block are removed from the sheath and insert. The cylinder is flipped over, and the exposed microwells are loaded with embryos, as described in Fig. 2 . (4) The cylinder and agarose block are lowered into the glass-bottom dish, and live-imaging medium is added in the cylinder. Right: Schematic of embryo in Configuration 3.

    Article Snippet: When making the microwells, agarose (Bio-Rad #1613101) was dissolved at 1.5% weight/volume (w/v) in distilled water (or 2% for firmer molds).

    Techniques: Imaging, Blocking Assay

    OMMAwell configurations for top-loaded microwells. (A) Configuration 1: Top loaded microwells for injecting or imaging with an upright microscope. (1) The mold insert (green) is inverted and connected to the slide (purple), which is placed into the upright platform (pink). (2) After the desired height is chosen, the pin (orange) is inserted. (3) Molten agarose is poured into a plastic petri dish and the mold assembly is lowered into it. (4) After the agarose sets, the mold insert is removed and eggs are placed into the wells, either individually with forceps or many at once by transferring the eggs in water with a cut plastic pipette. Excess water is removed by pipet and then wicked away with piece of lint-free lens paper. Then, 40–100 µl of molten low-melt agarose, kept at 42°C, is added to the wells to hold the eggs in place. Embryo positions are adjusted with plastic forceps. (5) When the low-melt agarose sets, the live-imaging medium is added to the dish. Right: Schematic of embryo in Configuration 1. (B) Configuration 2: Top loaded microwells for imaging with an inverted microscope. (1) 700 µl of agarose is pipetted into the middle of the glass-bottom dish. The insert and slide are lowered onto it, taking care not to trap bubbles. (2) Agarose sets, and then the insert and slide are gently removed. (3) Embryos are loaded into microwells, as described above. (4) Live-imaging medium is added. Right: Schematic of embryo in Configuration 2.

    Journal: Biology Open

    Article Title: High-throughput live-imaging of embryos in microwell arrays using a modular specimen mounting system

    doi: 10.1242/bio.031260

    Figure Lengend Snippet: OMMAwell configurations for top-loaded microwells. (A) Configuration 1: Top loaded microwells for injecting or imaging with an upright microscope. (1) The mold insert (green) is inverted and connected to the slide (purple), which is placed into the upright platform (pink). (2) After the desired height is chosen, the pin (orange) is inserted. (3) Molten agarose is poured into a plastic petri dish and the mold assembly is lowered into it. (4) After the agarose sets, the mold insert is removed and eggs are placed into the wells, either individually with forceps or many at once by transferring the eggs in water with a cut plastic pipette. Excess water is removed by pipet and then wicked away with piece of lint-free lens paper. Then, 40–100 µl of molten low-melt agarose, kept at 42°C, is added to the wells to hold the eggs in place. Embryo positions are adjusted with plastic forceps. (5) When the low-melt agarose sets, the live-imaging medium is added to the dish. Right: Schematic of embryo in Configuration 1. (B) Configuration 2: Top loaded microwells for imaging with an inverted microscope. (1) 700 µl of agarose is pipetted into the middle of the glass-bottom dish. The insert and slide are lowered onto it, taking care not to trap bubbles. (2) Agarose sets, and then the insert and slide are gently removed. (3) Embryos are loaded into microwells, as described above. (4) Live-imaging medium is added. Right: Schematic of embryo in Configuration 2.

    Article Snippet: When making the microwells, agarose (Bio-Rad #1613101) was dissolved at 1.5% weight/volume (w/v) in distilled water (or 2% for firmer molds).

    Techniques: Imaging, Microscopy, Transferring, Inverted Microscopy

    Overview of serum chemokines, cytokines and growth factors of the atrial fibrillation patient compared to control groups. Serum biomarkers: CXCL8 (IL-8); CXCL10 (IP-10); CCL11 (Eotaxin); CCL3 (MIP-1α); CCL4 (MIP-1β); CCL2 (MCP-1); CCL-5 (RANTES); IL1-β, IL-6, TNF-α, IL-12; IFN-γ, IL-17; IL-1Ra (IL-1 receptor antagonist); IL-2; IL-4; IL-5; IL-7; IL-9; IL-10; IL-13; IL-15; FGF-basic; PDGF; VEGF; G-CSF and GM-CSF were measured by high performance microbeads 27-plex assay, as described in materials and methods, with the 95%CI represented by gray boxes. Black arrows represent an increase (up) or decrease (down) in the patient’s biomarkers level in comparison to control groups; a black square represents biomarkers levels at the 95%CI.  a  Biomarkers’ levels: case vs. gender matching non-infected heath controls; ( b ) Biomarkers’ levels: case vs. gender matching ZIKV-infected patients

    Journal: Virology Journal

    Article Title: Atrial fibrillation in a patient with Zika virus infection

    doi: 10.1186/s12985-018-0938-2

    Figure Lengend Snippet: Overview of serum chemokines, cytokines and growth factors of the atrial fibrillation patient compared to control groups. Serum biomarkers: CXCL8 (IL-8); CXCL10 (IP-10); CCL11 (Eotaxin); CCL3 (MIP-1α); CCL4 (MIP-1β); CCL2 (MCP-1); CCL-5 (RANTES); IL1-β, IL-6, TNF-α, IL-12; IFN-γ, IL-17; IL-1Ra (IL-1 receptor antagonist); IL-2; IL-4; IL-5; IL-7; IL-9; IL-10; IL-13; IL-15; FGF-basic; PDGF; VEGF; G-CSF and GM-CSF were measured by high performance microbeads 27-plex assay, as described in materials and methods, with the 95%CI represented by gray boxes. Black arrows represent an increase (up) or decrease (down) in the patient’s biomarkers level in comparison to control groups; a black square represents biomarkers levels at the 95%CI. a Biomarkers’ levels: case vs. gender matching non-infected heath controls; ( b ) Biomarkers’ levels: case vs. gender matching ZIKV-infected patients

    Article Snippet: High-performance microbeads 27-plex assay (Bio-Rad, Hercules, CA, USA) was employed for detection and quantification of multiple targets, including: CXCL8 (IL-8); CXCL10 (IP-10); CCL11 (Eotaxin); CCL3 (MIP-1α); CCL4 (MIP-1β); CCL2 (MCP-1); CCL-5 (RANTES); IL1- β, IL-6, TNF- α, IL-12; IFN- γ, IL-17; IL-1Ra (IL-1 receptor antagonist); IL-2; IL-4; IL-5; IL-7; IL-9; IL-10; IL-13; IL-15; FGF-basic; PDGF; VEGF; G-CSF and GM-CSF.

    Techniques: Plex Assay, Infection

    A MR100-based assay can differentiate between prion-infected and normal brain homogenates without proteinase K digestion. a Schematic description of the RCA protocol to test brain homogenates without PK digestion. Brain tissues were freshly homogenized in microbead-containing tubes. Normal brain homogenates (NBH) or prion-infected brain homogenates (IBH) were incubated with MR100 for 1 h, at room temperature, leading to a precipitation of PrP isoforms. After a short centrifugation step, the pellet with MR100 (orange tube) concentrates PrP isoforms, whereas no pellet is detectable with DMSO. b Comparison of DMSO, P30 and MR100 precipitation capabilities using the RCA protocol. Fifty μL of 10 % 22 L infected brain homogenates were diluted in 300 μL of PBS/2 % sarcosyl and incubated using either 1.5 mM of P30 or MR100 or an equivalent volume of the solvent alone (DMSO), at room temperature for 1 h. Then, samples were centrifuged at 8000 g for 5 min. Supernatants (S) were collected and 30 μL of each supernatant was mixed with an equivalent volume of 2X loading buffer. Pellets (P) were resuspended in 30 μL PBS/2 % Sarcosyl, and mixed with an equal volume of 2X loading buffer. Thirty microliters of each sample were loaded on 12 % Bis-Tris gels (Criterion, Biorad) and immunoblotting was carried out with the SAF mix according to standard procedures [ 16 ]. The samples (S/P) were analyzed by western blotting using SAF mix anti-PrP antibodies. c Comparison of infected versus non-infected brain homogenates processed with the RCA protocol. Fifty microliters of 10 % freshly homogenized brain tissues from normal (NBH) or 22 L prion-infected (IBH) mice were processed according to the RCA protocol described in A and B. Thirty microliters of supernatant (S) or pellet (P) were loaded on 12 % Bis-Tris gels (Criterion, Biorad) and immunoblotting was carried out with the SAF mix as described above. Molecular masses (20–75 kDa) are indicated on the left side of the panels

    Journal: Molecular Neurodegeneration

    Article Title: A Fluorescent Oligothiophene-Bis-Triazine ligand interacts with PrP fibrils and detects SDS-resistant oligomers in human prion diseases

    doi: 10.1186/s13024-016-0074-7

    Figure Lengend Snippet: A MR100-based assay can differentiate between prion-infected and normal brain homogenates without proteinase K digestion. a Schematic description of the RCA protocol to test brain homogenates without PK digestion. Brain tissues were freshly homogenized in microbead-containing tubes. Normal brain homogenates (NBH) or prion-infected brain homogenates (IBH) were incubated with MR100 for 1 h, at room temperature, leading to a precipitation of PrP isoforms. After a short centrifugation step, the pellet with MR100 (orange tube) concentrates PrP isoforms, whereas no pellet is detectable with DMSO. b Comparison of DMSO, P30 and MR100 precipitation capabilities using the RCA protocol. Fifty μL of 10 % 22 L infected brain homogenates were diluted in 300 μL of PBS/2 % sarcosyl and incubated using either 1.5 mM of P30 or MR100 or an equivalent volume of the solvent alone (DMSO), at room temperature for 1 h. Then, samples were centrifuged at 8000 g for 5 min. Supernatants (S) were collected and 30 μL of each supernatant was mixed with an equivalent volume of 2X loading buffer. Pellets (P) were resuspended in 30 μL PBS/2 % Sarcosyl, and mixed with an equal volume of 2X loading buffer. Thirty microliters of each sample were loaded on 12 % Bis-Tris gels (Criterion, Biorad) and immunoblotting was carried out with the SAF mix according to standard procedures [ 16 ]. The samples (S/P) were analyzed by western blotting using SAF mix anti-PrP antibodies. c Comparison of infected versus non-infected brain homogenates processed with the RCA protocol. Fifty microliters of 10 % freshly homogenized brain tissues from normal (NBH) or 22 L prion-infected (IBH) mice were processed according to the RCA protocol described in A and B. Thirty microliters of supernatant (S) or pellet (P) were loaded on 12 % Bis-Tris gels (Criterion, Biorad) and immunoblotting was carried out with the SAF mix as described above. Molecular masses (20–75 kDa) are indicated on the left side of the panels

    Article Snippet: Brain tissues from terminally sick mice (infected with 22 L prions) and hamsters at various stages of disease (infected with the 263 K strain) were homogenized in 10 % (w/v) PBS using microbead-containing tubes and a Ribolysor apparatus (Biorad, Marnes la Coquette, France).

    Techniques: Infection, Incubation, Centrifugation, Western Blot, Mouse Assay

    Biochemical properties of SVmab and rSVmab. A Binding activity of SVmab and rSVmab in ELISA. hVSD II (1 μg) was coated on the microwell plate in 0.05 mmol/L carconate-bicarbonate buffer, pH 9.6. Anti-FLAG antibody was used as a negative control. All ELISA measurements were performed in triplicate. Data are mean ± SEM. B Antibody binding in ELISA. SVmab (66 nmol/L), SVmab (first batch, 66 nmol/L), and CTmab (66 nmol/L) were used. Anti-FLAG antibody (266 nmol/L) was added as a positive control. All ELISA measurements were performed in triplicate. Data are mean ± SEM. C SDS-PAGE (left) and western blots (right) of SVmab (2 μg) and rSVmab (2 μg) under a non-reducing condition. The gel (left) was stained with Coomassie blue. D SDS-PAGE of SVmab (1 μg) and rSVmab (1 μg) under a reducing condition. The gel was stained with Coomassie blue.

    Journal: Neuroscience Bulletin

    Article Title: Differential Inhibition of Nav1.7 and Neuropathic Pain by Hybridoma-Produced and Recombinant Monoclonal Antibodies that Target Nav1.7

    doi: 10.1007/s12264-018-0203-0

    Figure Lengend Snippet: Biochemical properties of SVmab and rSVmab. A Binding activity of SVmab and rSVmab in ELISA. hVSD II (1 μg) was coated on the microwell plate in 0.05 mmol/L carconate-bicarbonate buffer, pH 9.6. Anti-FLAG antibody was used as a negative control. All ELISA measurements were performed in triplicate. Data are mean ± SEM. B Antibody binding in ELISA. SVmab (66 nmol/L), SVmab (first batch, 66 nmol/L), and CTmab (66 nmol/L) were used. Anti-FLAG antibody (266 nmol/L) was added as a positive control. All ELISA measurements were performed in triplicate. Data are mean ± SEM. C SDS-PAGE (left) and western blots (right) of SVmab (2 μg) and rSVmab (2 μg) under a non-reducing condition. The gel (left) was stained with Coomassie blue. D SDS-PAGE of SVmab (1 μg) and rSVmab (1 μg) under a reducing condition. The gel was stained with Coomassie blue.

    Article Snippet: SFX-Insect cell medium was from Hyclone Laboratories (UT); n-dodecyl-β-D-maltopyranoside (DDM) for the human Nav1.7 VSDII purification was from Anatrace; carbonate-bicarbonate buffer was from Sigma and Tween-20 for ELISA was from Bio-Rad; ELISA microwell plates were from Nunc; anti-FLAG was from Sigma and HRP-conjugated secondary antibody was from Thermo Scientific; tetramethylbenzidine (TMB)-based substrate solution for an ELISA reaction was from KPL Inc. (MA); PVDF membranes and secondary antibody for western blotting of SVmab and rSVmab were from Bio-Rad and Li-Cor (IRDye® 800CW) (NE); paclitaxel was from Sigma.

    Techniques: Binding Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Negative Control, Positive Control, SDS Page, Western Blot, Staining