taq polymerase kit  (Thermo Fisher)


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    Name:
    DNA Polymerase I
    Description:
    DNA Polymerase I is a DNA polymerase with 5 →3 and 3 →5 exodeoxyribonuclease activities DNA Polymerase I also incorporates biotinylated nucleotides • Applications DNase I dependent nick translation second strand synthesis in cDNA cloning fill in of 5 overhangs• Source purified from E coli expressing the DNA Polymerase I gene on a plasmidPerformance and quality testing Endodeoxyribonuclease assay efficiency of DNase I dependent nick translation determined Unit definition One unit incorporates 10 nmol of total deoxyribonucleotide into acid precipitable material in 30 min at 37°C using template primer Unit reaction conditions 50 mM potassium phosphate pH 7 0 6 7 mM MgCl2 1 mM 2 mercaptoethanol 80 µg mL template primer 32 µM dTTP 69 nM 3H dTTP and enzyme in 100 µL for 30 min at 37°C
    Catalog Number:
    18010017
    Price:
    None
    Category:
    Proteins Enzymes Peptides
    Applications:
    Cloning|Restriction Enzyme Cloning
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    Structured Review

    Thermo Fisher taq polymerase kit
    DNA Polymerase I is a DNA polymerase with 5 →3 and 3 →5 exodeoxyribonuclease activities DNA Polymerase I also incorporates biotinylated nucleotides • Applications DNase I dependent nick translation second strand synthesis in cDNA cloning fill in of 5 overhangs• Source purified from E coli expressing the DNA Polymerase I gene on a plasmidPerformance and quality testing Endodeoxyribonuclease assay efficiency of DNase I dependent nick translation determined Unit definition One unit incorporates 10 nmol of total deoxyribonucleotide into acid precipitable material in 30 min at 37°C using template primer Unit reaction conditions 50 mM potassium phosphate pH 7 0 6 7 mM MgCl2 1 mM 2 mercaptoethanol 80 µg mL template primer 32 µM dTTP 69 nM 3H dTTP and enzyme in 100 µL for 30 min at 37°C
    https://www.bioz.com/result/taq polymerase kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    taq polymerase kit - by Bioz Stars, 2021-06
    99/100 stars

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    Related Articles

    Luciferase:

    Article Title: Cysteine Dioxygenase 1 Is a Tumor Suppressor Gene Silenced by Promoter Methylation in Multiple Human Cancers
    Article Snippet: .. Construction of Luciferase Vectors and Reporter Assay Potential CDO1 promoter regions upstream of the transcription start site (TSS) (−1100 and −430 bp to +104 bp) were prepared by PCR using pfx DNA polymerase (Invitrogen). ..

    Reporter Assay:

    Article Title: Cysteine Dioxygenase 1 Is a Tumor Suppressor Gene Silenced by Promoter Methylation in Multiple Human Cancers
    Article Snippet: .. Construction of Luciferase Vectors and Reporter Assay Potential CDO1 promoter regions upstream of the transcription start site (TSS) (−1100 and −430 bp to +104 bp) were prepared by PCR using pfx DNA polymerase (Invitrogen). ..

    Polymerase Chain Reaction:

    Article Title: Cysteine Dioxygenase 1 Is a Tumor Suppressor Gene Silenced by Promoter Methylation in Multiple Human Cancers
    Article Snippet: .. Construction of Luciferase Vectors and Reporter Assay Potential CDO1 promoter regions upstream of the transcription start site (TSS) (−1100 and −430 bp to +104 bp) were prepared by PCR using pfx DNA polymerase (Invitrogen). ..

    Article Title: Reconstituting ParA/ParB-mediated transport of DNA cargo
    Article Snippet: Restriction Digest Buffer #3 (New England Biolabs) Restriction Digest Buffer #4 (New England Biolabs) DNAPI Buffer: 50 mM Tris–HCl (pH 7.8), 5 mM MgCl2 , 10 mM 2-mercaptoethanol, 10 μg/mL of bovine serum albumin T4 Ligase Buffer (New England Biolabs; Cat. # M0202S) Tris–acetate–EDTA Buffer: 40 mM Tris, 20 mM Acetate, 1 mM EDTA TE Buffer: 10 mM Tris (pH 8), 1 mM EDTA .. pBR322 plasmid (4.4 Kb) pBR322:: sopC plasmid (4.7 Kb) Nt BspQI Nicking Enzyme (New England Biolabs, Cat. # R0644S) Phenol:CHCl3 :Isoamyl Alcohol (UltraPure 25:24:1 v/v) Ethanol Deoxyribonucleotide (dNTP) mix (Life Technologies; Cat. # 18427–013) dCTP (Life Technologies; Cat. # 18253–013) Alexa Fluor 647–12-OBEA-dCTP (Life Technologies; Cat. # ) DNA Polymerase I (Life Technologies, Cat. # 18010–025) SYBR Gold DNA Stain (Life Technologies, Cat. # S-11494) Illustra MicroSpin S-400HR columns (GE Healthcare, Cat # 27–5140-01) QIAquick polymerase chain reaction (PCR) Purification Kit (Qiagen, Cat. # 28104) Ethidium Bromide 400 U/μL T4 Ligase (New England Biolabs, Cat. # M0202S) .. Water Bath Vortex Table top centrifuge Nanodrop Spectrophotometer Typhoon Imager (GE Healthcare).

    Article Title: SETD2 loss-of-function promotes renal cancer branched evolution through replication stress and impaired DNA repair
    Article Snippet: .. PCR reactions were performed using two different DNA polymerase enzymes to exclude any enzyme-specific effects, and the products were subsequently subjected to fragment analysis using the ABI3130XL Genetic Analyzer (Life Technologies Ltd, Paisley, UK) .. Thymidine block Cells were treated with 2 mM thymidine for 18 h, washed with phosphate-buffered saline and released into complete media for 9 h. Cells were subsequently treated with a second thymidine block for a further 18 h. Cells were washed, released into complete media and harvested at the indicated time points.

    Article Title: A LysR-family transcriptional regulator required for virulence in Brucella abortus is highly conserved among the α-proteobacteria
    Article Snippet: .. An approximately 1 kb fragment of the upstream region of the gene was amplified by PCR using the primers BAB1_1517-Up-For and BAB1_1517-Up-Rev, genomic DNA from B. abortus 2308 as a template, and Platinum® Pfx DNA Polymerase (Invitrogen, Carlsbad, CA, USA). .. Likewise, an approximately 1 kb fragment of the downstream region of the gene was amplified by PCR using the primers BAB1_1517-Down-For and BAB1_1517-Down-Rev.

    Plasmid Preparation:

    Article Title: Reconstituting ParA/ParB-mediated transport of DNA cargo
    Article Snippet: Restriction Digest Buffer #3 (New England Biolabs) Restriction Digest Buffer #4 (New England Biolabs) DNAPI Buffer: 50 mM Tris–HCl (pH 7.8), 5 mM MgCl2 , 10 mM 2-mercaptoethanol, 10 μg/mL of bovine serum albumin T4 Ligase Buffer (New England Biolabs; Cat. # M0202S) Tris–acetate–EDTA Buffer: 40 mM Tris, 20 mM Acetate, 1 mM EDTA TE Buffer: 10 mM Tris (pH 8), 1 mM EDTA .. pBR322 plasmid (4.4 Kb) pBR322:: sopC plasmid (4.7 Kb) Nt BspQI Nicking Enzyme (New England Biolabs, Cat. # R0644S) Phenol:CHCl3 :Isoamyl Alcohol (UltraPure 25:24:1 v/v) Ethanol Deoxyribonucleotide (dNTP) mix (Life Technologies; Cat. # 18427–013) dCTP (Life Technologies; Cat. # 18253–013) Alexa Fluor 647–12-OBEA-dCTP (Life Technologies; Cat. # ) DNA Polymerase I (Life Technologies, Cat. # 18010–025) SYBR Gold DNA Stain (Life Technologies, Cat. # S-11494) Illustra MicroSpin S-400HR columns (GE Healthcare, Cat # 27–5140-01) QIAquick polymerase chain reaction (PCR) Purification Kit (Qiagen, Cat. # 28104) Ethidium Bromide 400 U/μL T4 Ligase (New England Biolabs, Cat. # M0202S) .. Water Bath Vortex Table top centrifuge Nanodrop Spectrophotometer Typhoon Imager (GE Healthcare).

    Article Title: A polyglutamine domain is required for de novo CIZ1 assembly formation at the inactive X chromosome
    Article Snippet: Purified proteins were supplemented to 9% v/v (final concentration) with sterile glycerol, aliquoted and snap frozen in nuclease-free cryo tubes (Nunc™) in liquid nitrogen and stored at −80°C. .. In vitro transcription of DIG labelled probes DNA templates used for in vitro transcription of mouse Xist RNA were amplified from sequence verified plasmid pCMV-Xist-PA ( ) (Addgene 26760), containing the murine Xist gene using high-fidelity Platinum® pfx DNA polymerase (Invitrogen™). .. PCR primers used for the amplifications contained T7 promoter sequences, were designed with SnapGene (GraphPad Software), and are shown in Supplemental Table 2.

    Staining:

    Article Title: Reconstituting ParA/ParB-mediated transport of DNA cargo
    Article Snippet: Restriction Digest Buffer #3 (New England Biolabs) Restriction Digest Buffer #4 (New England Biolabs) DNAPI Buffer: 50 mM Tris–HCl (pH 7.8), 5 mM MgCl2 , 10 mM 2-mercaptoethanol, 10 μg/mL of bovine serum albumin T4 Ligase Buffer (New England Biolabs; Cat. # M0202S) Tris–acetate–EDTA Buffer: 40 mM Tris, 20 mM Acetate, 1 mM EDTA TE Buffer: 10 mM Tris (pH 8), 1 mM EDTA .. pBR322 plasmid (4.4 Kb) pBR322:: sopC plasmid (4.7 Kb) Nt BspQI Nicking Enzyme (New England Biolabs, Cat. # R0644S) Phenol:CHCl3 :Isoamyl Alcohol (UltraPure 25:24:1 v/v) Ethanol Deoxyribonucleotide (dNTP) mix (Life Technologies; Cat. # 18427–013) dCTP (Life Technologies; Cat. # 18253–013) Alexa Fluor 647–12-OBEA-dCTP (Life Technologies; Cat. # ) DNA Polymerase I (Life Technologies, Cat. # 18010–025) SYBR Gold DNA Stain (Life Technologies, Cat. # S-11494) Illustra MicroSpin S-400HR columns (GE Healthcare, Cat # 27–5140-01) QIAquick polymerase chain reaction (PCR) Purification Kit (Qiagen, Cat. # 28104) Ethidium Bromide 400 U/μL T4 Ligase (New England Biolabs, Cat. # M0202S) .. Water Bath Vortex Table top centrifuge Nanodrop Spectrophotometer Typhoon Imager (GE Healthcare).

    Purification:

    Article Title: Reconstituting ParA/ParB-mediated transport of DNA cargo
    Article Snippet: Restriction Digest Buffer #3 (New England Biolabs) Restriction Digest Buffer #4 (New England Biolabs) DNAPI Buffer: 50 mM Tris–HCl (pH 7.8), 5 mM MgCl2 , 10 mM 2-mercaptoethanol, 10 μg/mL of bovine serum albumin T4 Ligase Buffer (New England Biolabs; Cat. # M0202S) Tris–acetate–EDTA Buffer: 40 mM Tris, 20 mM Acetate, 1 mM EDTA TE Buffer: 10 mM Tris (pH 8), 1 mM EDTA .. pBR322 plasmid (4.4 Kb) pBR322:: sopC plasmid (4.7 Kb) Nt BspQI Nicking Enzyme (New England Biolabs, Cat. # R0644S) Phenol:CHCl3 :Isoamyl Alcohol (UltraPure 25:24:1 v/v) Ethanol Deoxyribonucleotide (dNTP) mix (Life Technologies; Cat. # 18427–013) dCTP (Life Technologies; Cat. # 18253–013) Alexa Fluor 647–12-OBEA-dCTP (Life Technologies; Cat. # ) DNA Polymerase I (Life Technologies, Cat. # 18010–025) SYBR Gold DNA Stain (Life Technologies, Cat. # S-11494) Illustra MicroSpin S-400HR columns (GE Healthcare, Cat # 27–5140-01) QIAquick polymerase chain reaction (PCR) Purification Kit (Qiagen, Cat. # 28104) Ethidium Bromide 400 U/μL T4 Ligase (New England Biolabs, Cat. # M0202S) .. Water Bath Vortex Table top centrifuge Nanodrop Spectrophotometer Typhoon Imager (GE Healthcare).

    Labeling:

    Article Title: Assigning functions to genes: identification of S-phase expressed genes in Leishmania major based on post-transcriptional control elements
    Article Snippet: .. DNA probes were labeled using [α-32 P]-dCTP, using random primers (Amersham Bioscience) and DNA polymerase I Klenow fragment (Fermentas), following the manufacturer's recommended protocol. .. Northern blot hybridization analysis Total cell RNA was isolated from samples of synchronous L.major cells at 1 h intervals after the removal of hydroxyurea, using TRI REAGENT™ (Molecular Research Center Inc.), following the manufacturer's procedure at a ratio of 2.5–5 × 108 L.major cells/ml TRI REAGENT™.

    In Vitro:

    Article Title: A polyglutamine domain is required for de novo CIZ1 assembly formation at the inactive X chromosome
    Article Snippet: Purified proteins were supplemented to 9% v/v (final concentration) with sterile glycerol, aliquoted and snap frozen in nuclease-free cryo tubes (Nunc™) in liquid nitrogen and stored at −80°C. .. In vitro transcription of DIG labelled probes DNA templates used for in vitro transcription of mouse Xist RNA were amplified from sequence verified plasmid pCMV-Xist-PA ( ) (Addgene 26760), containing the murine Xist gene using high-fidelity Platinum® pfx DNA polymerase (Invitrogen™). .. PCR primers used for the amplifications contained T7 promoter sequences, were designed with SnapGene (GraphPad Software), and are shown in Supplemental Table 2.

    Amplification:

    Article Title: A polyglutamine domain is required for de novo CIZ1 assembly formation at the inactive X chromosome
    Article Snippet: Purified proteins were supplemented to 9% v/v (final concentration) with sterile glycerol, aliquoted and snap frozen in nuclease-free cryo tubes (Nunc™) in liquid nitrogen and stored at −80°C. .. In vitro transcription of DIG labelled probes DNA templates used for in vitro transcription of mouse Xist RNA were amplified from sequence verified plasmid pCMV-Xist-PA ( ) (Addgene 26760), containing the murine Xist gene using high-fidelity Platinum® pfx DNA polymerase (Invitrogen™). .. PCR primers used for the amplifications contained T7 promoter sequences, were designed with SnapGene (GraphPad Software), and are shown in Supplemental Table 2.

    Article Title: A LysR-family transcriptional regulator required for virulence in Brucella abortus is highly conserved among the α-proteobacteria
    Article Snippet: .. An approximately 1 kb fragment of the upstream region of the gene was amplified by PCR using the primers BAB1_1517-Up-For and BAB1_1517-Up-Rev, genomic DNA from B. abortus 2308 as a template, and Platinum® Pfx DNA Polymerase (Invitrogen, Carlsbad, CA, USA). .. Likewise, an approximately 1 kb fragment of the downstream region of the gene was amplified by PCR using the primers BAB1_1517-Down-For and BAB1_1517-Down-Rev.

    Sequencing:

    Article Title: A polyglutamine domain is required for de novo CIZ1 assembly formation at the inactive X chromosome
    Article Snippet: Purified proteins were supplemented to 9% v/v (final concentration) with sterile glycerol, aliquoted and snap frozen in nuclease-free cryo tubes (Nunc™) in liquid nitrogen and stored at −80°C. .. In vitro transcription of DIG labelled probes DNA templates used for in vitro transcription of mouse Xist RNA were amplified from sequence verified plasmid pCMV-Xist-PA ( ) (Addgene 26760), containing the murine Xist gene using high-fidelity Platinum® pfx DNA polymerase (Invitrogen™). .. PCR primers used for the amplifications contained T7 promoter sequences, were designed with SnapGene (GraphPad Software), and are shown in Supplemental Table 2.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Promoter Hypermethylation of CHODL Contributes to Carcinogenesis and Indicates Poor Survival in Patients with Early-stage Colorectal Cancer
    Article Snippet: PCR was performed to detect relative CHODL expression. .. Semiquantitative RT-PCR was performed using Hot Start DNA Polymerase (Invitrogen). ..

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    Thermo Fisher superscript iii one step rt pcr taq high fidelity
    Seckel mouse spermatocytes exhibit more γH2AX patches than wild-type cells. a <t>RT-PCR</t> using primers that anneal to Atr exons 8 and 10. Atr S/S testis exhibits two main RT-PCR products, one corresponding to full-length Atr (FL Atr , 477 bp), which is substantially reduced, and another corresponding to Atr lacking exon 9 (ΔE9 Atr . RT-PCR for β Actin is also provided as a control. b Percentage of cells exhibiting ATR staining extended to X and Y chromatin or confined to the X and Y chromosome axes in Atr +/+ and Atr S/S ( N = 284 and N = 282, respectively). Columns and lines indicate the mean and standard deviation (SD) from analysis performed on <t>three</t> wild-type and three mutant mice. Images show ATR straining on the sex body in Atr +/+ and Atr S/S pachytene spermatocytes. Images were captured with the same exposure time. Scale bar = 2 μm. c Quantification of the intensity of γH2AX staining on the sex body in arbitrary units (a.u.). Horizontal black lines denote the means. Images show representative sex bodies from Atr +/+ and Atr S/S pachytene spermatocytes immunostained against γH2AX. Images were captured with the same exposure time. Scale bar = 2 μm. Representative images of Atr +/+ ( d ) and Atr S/S ( e ) spermatocytes at different stages of meiotic prophase immunostained against SYCP3, γH2AX, and H1T. Scale bars = 10 μm. Numbers represent the mean ± SD of γH2AX patches found in each stage and genotype. N . Arrowheads indicate examples of aberrant autosomal γH2AX patches in Atr S/S spermatocytes. The differences between controls and Atr S/S for γH2AX patch numbers at late pachynema and diplonema were statistically significant ( p = 0.0001 and p = 0.0007, respectively, t test)
    Superscript Iii One Step Rt Pcr Taq High Fidelity, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii one step rt pcr taq high fidelity/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    99
    Thermo Fisher cells direct qrt pcr kit
    Fluidigm <t>qRT–PCR</t> analysis of 23GFP + (green) and 23GFP – (blue) VE-Cadh + Ter119 – CD45 – CD41 – ECs and/or VE-Cadh + Ter119 – CD45 – CD41 + HPCs (grey) from E8.5 (5–9 sp), 9.25 (18-22 sp), and 10.5
    Cells Direct Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cells direct qrt pcr kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    99
    Thermo Fisher superscript iii platinum sybr green one step qrt pcr kit
    Clinical validation of ai-LAMP. ( A – C ) Comparative sample positivity between LAMP and RdRP <t>qRT-PCR</t> ( A ), LAMP and N qRT-PCR ( B ), LAMP and CUM qRT-PCR results ( C ). ( D ) The heatmap indicates the relative positive and negative samples among <t>three</t> assays. ( E ) Linearity chart comparing the LAMP positive/negative samples and their detection based on the RdRP gene-based qRT-PCR. ( F ) Linearity chart comparing the LAMP positive/negative samples and their detection based on the N gene-based qRT-PCR. ( G ) Naked eye detection of the first 96 samples out of the total 199 patients’ samples were processed. ( H ) Recovery Ct values of the miRNA spiked before RNA extraction.
    Superscript Iii Platinum Sybr Green One Step Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii platinum sybr green one step qrt pcr kit/product/Thermo Fisher
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    Image Search Results


    Seckel mouse spermatocytes exhibit more γH2AX patches than wild-type cells. a RT-PCR using primers that anneal to Atr exons 8 and 10. Atr S/S testis exhibits two main RT-PCR products, one corresponding to full-length Atr (FL Atr , 477 bp), which is substantially reduced, and another corresponding to Atr lacking exon 9 (ΔE9 Atr . RT-PCR for β Actin is also provided as a control. b Percentage of cells exhibiting ATR staining extended to X and Y chromatin or confined to the X and Y chromosome axes in Atr +/+ and Atr S/S ( N = 284 and N = 282, respectively). Columns and lines indicate the mean and standard deviation (SD) from analysis performed on three wild-type and three mutant mice. Images show ATR straining on the sex body in Atr +/+ and Atr S/S pachytene spermatocytes. Images were captured with the same exposure time. Scale bar = 2 μm. c Quantification of the intensity of γH2AX staining on the sex body in arbitrary units (a.u.). Horizontal black lines denote the means. Images show representative sex bodies from Atr +/+ and Atr S/S pachytene spermatocytes immunostained against γH2AX. Images were captured with the same exposure time. Scale bar = 2 μm. Representative images of Atr +/+ ( d ) and Atr S/S ( e ) spermatocytes at different stages of meiotic prophase immunostained against SYCP3, γH2AX, and H1T. Scale bars = 10 μm. Numbers represent the mean ± SD of γH2AX patches found in each stage and genotype. N . Arrowheads indicate examples of aberrant autosomal γH2AX patches in Atr S/S spermatocytes. The differences between controls and Atr S/S for γH2AX patch numbers at late pachynema and diplonema were statistically significant ( p = 0.0001 and p = 0.0007, respectively, t test)

    Journal: Nature Communications

    Article Title: ATR is required to complete meiotic recombination in mice

    doi: 10.1038/s41467-018-04851-z

    Figure Lengend Snippet: Seckel mouse spermatocytes exhibit more γH2AX patches than wild-type cells. a RT-PCR using primers that anneal to Atr exons 8 and 10. Atr S/S testis exhibits two main RT-PCR products, one corresponding to full-length Atr (FL Atr , 477 bp), which is substantially reduced, and another corresponding to Atr lacking exon 9 (ΔE9 Atr . RT-PCR for β Actin is also provided as a control. b Percentage of cells exhibiting ATR staining extended to X and Y chromatin or confined to the X and Y chromosome axes in Atr +/+ and Atr S/S ( N = 284 and N = 282, respectively). Columns and lines indicate the mean and standard deviation (SD) from analysis performed on three wild-type and three mutant mice. Images show ATR straining on the sex body in Atr +/+ and Atr S/S pachytene spermatocytes. Images were captured with the same exposure time. Scale bar = 2 μm. c Quantification of the intensity of γH2AX staining on the sex body in arbitrary units (a.u.). Horizontal black lines denote the means. Images show representative sex bodies from Atr +/+ and Atr S/S pachytene spermatocytes immunostained against γH2AX. Images were captured with the same exposure time. Scale bar = 2 μm. Representative images of Atr +/+ ( d ) and Atr S/S ( e ) spermatocytes at different stages of meiotic prophase immunostained against SYCP3, γH2AX, and H1T. Scale bars = 10 μm. Numbers represent the mean ± SD of γH2AX patches found in each stage and genotype. N . Arrowheads indicate examples of aberrant autosomal γH2AX patches in Atr S/S spermatocytes. The differences between controls and Atr S/S for γH2AX patch numbers at late pachynema and diplonema were statistically significant ( p = 0.0001 and p = 0.0007, respectively, t test)

    Article Snippet: The SuperScript III One-Step RT-PCR Taq High Fidelity (Invitrogen) was used to synthesize cDNA and amplify Atr transcripts using previously designed primers .

    Techniques: Reverse Transcription Polymerase Chain Reaction, Staining, Standard Deviation, Mutagenesis, Mouse Assay

    Simplified designs used to rescue infectious RNA viruses. The entire viral genome, flanked respectively at the 5’ and 3’ untranslated regions (UTRs) by the human cytomegalovirus promoter (pCMV) and the hepatitis delta ribozyme followed by the simian virus 40 polyadenylation signal (HDR/SV40pA) is schematized at the top of the figure. For each design, called A to E and the classical ISA design (named ‘ISA’), the number of overlapping subgenomic amplicons ranged from three to five (Left panel). These amplicons were amplified by PCR/RT-PCR, purified, pooled and then transfected into permissive cells. Each design was tested with a panel of viruses in mosquito (C6/36, U4.4) and mammalian (SW13, HEK-293) cell lines. After cell transfection and two serial passages, virus replication was demonstrated using a combination of several criteria (CPE, production of viral RNAs and infectious particles in cell supernatant medium). The percentage of success to recover a virus was calculated using the result of four independent experiments of each procedure and is reported (Right panel).

    Journal: PLoS ONE

    Article Title: Haiku: New paradigm for the reverse genetics of emerging RNA viruses

    doi: 10.1371/journal.pone.0193069

    Figure Lengend Snippet: Simplified designs used to rescue infectious RNA viruses. The entire viral genome, flanked respectively at the 5’ and 3’ untranslated regions (UTRs) by the human cytomegalovirus promoter (pCMV) and the hepatitis delta ribozyme followed by the simian virus 40 polyadenylation signal (HDR/SV40pA) is schematized at the top of the figure. For each design, called A to E and the classical ISA design (named ‘ISA’), the number of overlapping subgenomic amplicons ranged from three to five (Left panel). These amplicons were amplified by PCR/RT-PCR, purified, pooled and then transfected into permissive cells. Each design was tested with a panel of viruses in mosquito (C6/36, U4.4) and mammalian (SW13, HEK-293) cell lines. After cell transfection and two serial passages, virus replication was demonstrated using a combination of several criteria (CPE, production of viral RNAs and infectious particles in cell supernatant medium). The percentage of success to recover a virus was calculated using the result of four independent experiments of each procedure and is reported (Right panel).

    Article Snippet: When a RNA template was used (nucleic acid extract from cell supernatant medium or mice brain suspension filtrate), amplicons were generated by RT-PCR using the Superscript III One-Step RT-PCR Platinum Taq High Fidelity kit (Life Technologies) and a Biometra TProfessional Standard Gradient thermocycler as previously described [ ].

    Techniques: Amplification, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Purification, Transfection

    Fluidigm qRT–PCR analysis of 23GFP + (green) and 23GFP – (blue) VE-Cadh + Ter119 – CD45 – CD41 – ECs and/or VE-Cadh + Ter119 – CD45 – CD41 + HPCs (grey) from E8.5 (5–9 sp), 9.25 (18-22 sp), and 10.5

    Journal: Nature communications

    Article Title: Early dynamic fate changes in haemogenic endothelium characterized at the single-cell level

    doi: 10.1038/ncomms3924

    Figure Lengend Snippet: Fluidigm qRT–PCR analysis of 23GFP + (green) and 23GFP – (blue) VE-Cadh + Ter119 – CD45 – CD41 – ECs and/or VE-Cadh + Ter119 – CD45 – CD41 + HPCs (grey) from E8.5 (5–9 sp), 9.25 (18-22 sp), and 10.5

    Article Snippet: Cells were harvested directly into 10 μl RT-PreAmp Master Mix (consisting of 5 μl CellsDirect 2× Reaction Mix (Invitrogen), 2.5 μl 0.2× assay mix, 1.2 μl RT/Taq enzyme (Cells-Direct qRT–PCR kit, Invitrogen), 1.2 μl TE buffer (Invitrogen) and 0.1 μl SUPERase-In (Ambion)).

    Techniques: Quantitative RT-PCR

    Clinical validation of ai-LAMP. ( A – C ) Comparative sample positivity between LAMP and RdRP qRT-PCR ( A ), LAMP and N qRT-PCR ( B ), LAMP and CUM qRT-PCR results ( C ). ( D ) The heatmap indicates the relative positive and negative samples among three assays. ( E ) Linearity chart comparing the LAMP positive/negative samples and their detection based on the RdRP gene-based qRT-PCR. ( F ) Linearity chart comparing the LAMP positive/negative samples and their detection based on the N gene-based qRT-PCR. ( G ) Naked eye detection of the first 96 samples out of the total 199 patients’ samples were processed. ( H ) Recovery Ct values of the miRNA spiked before RNA extraction.

    Journal: Viruses

    Article Title: Artificial Intelligence-Assisted Loop Mediated Isothermal Amplification (AI-LAMP) for Rapid Detection of SARS-CoV-2

    doi: 10.3390/v12090972

    Figure Lengend Snippet: Clinical validation of ai-LAMP. ( A – C ) Comparative sample positivity between LAMP and RdRP qRT-PCR ( A ), LAMP and N qRT-PCR ( B ), LAMP and CUM qRT-PCR results ( C ). ( D ) The heatmap indicates the relative positive and negative samples among three assays. ( E ) Linearity chart comparing the LAMP positive/negative samples and their detection based on the RdRP gene-based qRT-PCR. ( F ) Linearity chart comparing the LAMP positive/negative samples and their detection based on the N gene-based qRT-PCR. ( G ) Naked eye detection of the first 96 samples out of the total 199 patients’ samples were processed. ( H ) Recovery Ct values of the miRNA spiked before RNA extraction.

    Article Snippet: The 25-µL PCR reaction consists of 12.5 µL 2X Reaction Mix, 0.2 µM of each primer and 0.1 µM probe, 0.5 µL of SuperScript® III RT/Platinum® Taq Mix, 5 µL of RNA sample and nuclear free water.

    Techniques: Quantitative RT-PCR, RNA Extraction