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Bio-Rad iscript cdna synthesis kit
Iscript Cdna Synthesis Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 3427 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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iscript cdna synthesis kit - by Bioz Stars, 2020-03
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DNA Extraction:

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: After centrifugation at 12000 × g for 10 minutes, the interphase was collected for DNA extraction while the RNA in the aqueous phase was precipitated with isopropanol, washed in 75% ethanol and then resuspended in 20 μ l RNAse-free water for storage at -80°C. .. Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions.

Nucleic Acid Electrophoresis:

Article Title: MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages
Article Snippet: Quantitative RT-PCR (Q-RT-PCR) RNA was extracted from pancreata using Trizol (Invitrogen) and quality checked by gel electrophoresis. .. Cells were incubated for the indicated times or overnight, after which time RNA was isolated using Trizol, then treated with DNAase1 (Invitrogen). cDNA was synthesized from 1 μg of RNA using iScript cDNA Synthesis Kit (BioRad).

Centrifugation:

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: After centrifugation at 12000 × g for 10 minutes, the interphase was collected for DNA extraction while the RNA in the aqueous phase was precipitated with isopropanol, washed in 75% ethanol and then resuspended in 20 μ l RNAse-free water for storage at -80°C. .. Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions.

Incubation:

Article Title: MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages
Article Snippet: .. Cells were incubated for the indicated times or overnight, after which time RNA was isolated using Trizol, then treated with DNAase1 (Invitrogen). cDNA was synthesized from 1 μg of RNA using iScript cDNA Synthesis Kit (BioRad). .. Q-RT-PCR was performed using SoFast EvagGreen Supermix (BioRad) with cDNA corresponding to 50ng of total RNA and 500nM primers in a 20μl volume.

RNA Sequencing Assay:

Article Title: Wnt-3a Induces Cytokine Release in Human Mast Cells
Article Snippet: .. Quantitative PCR (qPCR) and RNA Sequencing (RNAseq) For qPCR, RNA was extracted using an RNeasy Plus Mini Kit (Qiagen, Hilden, Germany), cDNA was prepared using an iScript cDNA synthesis kit (Bio-Rad), and qPCR was performed using TaqMan™ Gene Expression master mix (Thermo Fisher Scientific) on a CFX96 Real-Time System (Bio-Rad). .. The following primers were used: Hs02800695_m1 (HPRT1), Hs00174103_m1 (CXCL8), Hs04187715_m1 (CCL8), Hs00171147_m1 (CCL7), Hs01099660_g1 (CXCL5), Hs00234142_m1 (CCL3), and TaqMan Array Human WNT Pathway (all from Thermo Fisher Scientific).

Synthesized:

Article Title: Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating P38 Mitogen-Activated Protein Kinase Expression
Article Snippet: .. The cDNA was synthesized from 1 μg of RNA by applying to the Bio-Rad iScript cDNA synthesis kit (Bio-Rad). .. Quantitative polymerase chain reaction was performed on the MJ Opticon (Bio-Rad) system using iTaq Universal SYBR Green Supermix (Bio-Rad).

Article Title: Long-chain acyl-CoA synthetase 1 interacts with key proteins that activate and direct fatty acids into niche hepatic pathways
Article Snippet: .. Total RNA and DNA were simultaneously isolated from liver using 0.7 ml of TRIzol (ThermoFisher Scientific) per 200 mg of tissue. cDNA was synthesized using 1 μg of RNA template (iScript cDNA Synthesis kit, Bio-Rad), and 50 ng of cDNA was added to each qPCR with SYBR Green (iTaq Universal SYBR; Bio-Rad) and used to detect amplicons with primers specific to the gene of interest (S6) and quantified using a qPCR thermocycler (Bio-Rad). ..

Article Title: MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages
Article Snippet: .. Cells were incubated for the indicated times or overnight, after which time RNA was isolated using Trizol, then treated with DNAase1 (Invitrogen). cDNA was synthesized from 1 μg of RNA using iScript cDNA Synthesis Kit (BioRad). .. Q-RT-PCR was performed using SoFast EvagGreen Supermix (BioRad) with cDNA corresponding to 50ng of total RNA and 500nM primers in a 20μl volume.

Article Title: Improvement of epidermal covering on AEC patients with severe skin erosions by PRIMA-1MET/APR-246
Article Snippet: .. RNA was then extracted using RNEasy Mini kit (Qiagen), and cDNA was synthesized from 1 µg of RNA using iScript cDNA synthesis kit (Bio-Rad). .. Quantitative PCR was performed in triplicate using 2× SYBR Green PCR Master Mix (Absource Biotools).

Isolation:

Article Title: Restoring colistin sensitivity in colistin-resistant E. coli: Combinatorial use of MarR inhibitor with efflux pump inhibitor
Article Snippet: .. Total RNA from the bacterial strains were isolated using Aurum™ total RNA mini kit (Bio Rad). cDNA conversion was done using the iScript™ cDNA synthesis kit (Bio Rad). .. Both RNA and cDNA was quantified using Qubit fluorimeter (Invitrogen).

Article Title: The Ataxia telangiectasia-mutated and Rad3-related Protein Kinase Regulates Cellular Hydrogen Sulfide Concentrations
Article Snippet: Total cellular RNA was isolated from ATR and ATR-H cultured cells using Trizol according to the manufacturer’s instruction. .. One μg of RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-rad, 1708891).

Article Title: Endophytes alleviate the elevated CO2-dependent decrease in photosynthesis in rice, particularly under nitrogen limitation
Article Snippet: .. The isolated mRNAs were mixed with a master reaction mixture containing fluorescent dyes (iQ SyBR Green Supermix, Bio-Rad Laboratories) prior to cDNA synthesis using an iScript cDNA Synthesis Kit (Bio-Rad Laboratories) and were then loaded into a real-time qPCR thermocycler (Chromo 4 System, Bio-Rad Laboratories). .. The relative gene expression of rbcS was calculated according to .

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: Paragraph title: 2.3. RNA Isolation and Gene Expression ... Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions.

Article Title: Wnt-3a Induces Cytokine Release in Human Mast Cells
Article Snippet: Quantitative PCR (qPCR) and RNA Sequencing (RNAseq) For qPCR, RNA was extracted using an RNeasy Plus Mini Kit (Qiagen, Hilden, Germany), cDNA was prepared using an iScript cDNA synthesis kit (Bio-Rad), and qPCR was performed using TaqMan™ Gene Expression master mix (Thermo Fisher Scientific) on a CFX96 Real-Time System (Bio-Rad). .. For RNAseq, RNA was extracted using Arcturus PicoPure RNA Isolation Kit (Thermo Fisher Scientific).

Article Title: Long-chain acyl-CoA synthetase 1 interacts with key proteins that activate and direct fatty acids into niche hepatic pathways
Article Snippet: .. Total RNA and DNA were simultaneously isolated from liver using 0.7 ml of TRIzol (ThermoFisher Scientific) per 200 mg of tissue. cDNA was synthesized using 1 μg of RNA template (iScript cDNA Synthesis kit, Bio-Rad), and 50 ng of cDNA was added to each qPCR with SYBR Green (iTaq Universal SYBR; Bio-Rad) and used to detect amplicons with primers specific to the gene of interest (S6) and quantified using a qPCR thermocycler (Bio-Rad). ..

Article Title: MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages
Article Snippet: .. Cells were incubated for the indicated times or overnight, after which time RNA was isolated using Trizol, then treated with DNAase1 (Invitrogen). cDNA was synthesized from 1 μg of RNA using iScript cDNA Synthesis Kit (BioRad). .. Q-RT-PCR was performed using SoFast EvagGreen Supermix (BioRad) with cDNA corresponding to 50ng of total RNA and 500nM primers in a 20μl volume.

Sequencing:

Article Title: Restoring colistin sensitivity in colistin-resistant E. coli: Combinatorial use of MarR inhibitor with efflux pump inhibitor
Article Snippet: Paragraph title: Whole genome sequencing ... Total RNA from the bacterial strains were isolated using Aurum™ total RNA mini kit (Bio Rad). cDNA conversion was done using the iScript™ cDNA synthesis kit (Bio Rad).

Article Title: Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating P38 Mitogen-Activated Protein Kinase Expression
Article Snippet: The primers sequence are presented in . .. The cDNA was synthesized from 1 μg of RNA by applying to the Bio-Rad iScript cDNA synthesis kit (Bio-Rad).

RNA Extraction:

Article Title: Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
Article Snippet: Paragraph title: 2.5. Total RNA Extraction and Reverse Transcription ... Five hours after removal of bacteria, RNA was extracted directly from monolayers within the 12-well plates using the RNeasy Micro Kit (QIAGEN, Venlo, The Netherlands) according to the manufacturer’s instructions and converted to cDNA using the iSCRIPT cDNA Synthesis Kit (BioRad, Hercules, CA, USA).

Article Title: Long-chain acyl-CoA synthetase 1 interacts with key proteins that activate and direct fatty acids into niche hepatic pathways
Article Snippet: Paragraph title: RNA extraction and quantitative real-time PCR ... Total RNA and DNA were simultaneously isolated from liver using 0.7 ml of TRIzol (ThermoFisher Scientific) per 200 mg of tissue. cDNA was synthesized using 1 μg of RNA template (iScript cDNA Synthesis kit, Bio-Rad), and 50 ng of cDNA was added to each qPCR with SYBR Green (iTaq Universal SYBR; Bio-Rad) and used to detect amplicons with primers specific to the gene of interest (S6) and quantified using a qPCR thermocycler (Bio-Rad).

Cell Culture:

Article Title: The Ataxia telangiectasia-mutated and Rad3-related Protein Kinase Regulates Cellular Hydrogen Sulfide Concentrations
Article Snippet: Total cellular RNA was isolated from ATR and ATR-H cultured cells using Trizol according to the manufacturer’s instruction. .. One μg of RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-rad, 1708891).

Concentration Assay:

Article Title: Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
Article Snippet: Five hours after removal of bacteria, RNA was extracted directly from monolayers within the 12-well plates using the RNeasy Micro Kit (QIAGEN, Venlo, The Netherlands) according to the manufacturer’s instructions and converted to cDNA using the iSCRIPT cDNA Synthesis Kit (BioRad, Hercules, CA, USA). .. Quantitative real-time PCR (qPCR) Master-mix was prepared as follows: 5 µL Iq Sybr green Master-mix (Biorad), 1 µL of forward and reverse primers each (final concentration 100 nM), 1 µL of demineralized water and 2 µL of 1:20 diluted cDNA.

Article Title: Age-Related Changes in HAPLN1 Increase Lymphatic Permeability and Affect Routes of Melanoma Metastasis
Article Snippet: RNA concentration was measured using Nanodrop 2000 (Thermo Scientific). .. One microgram of RNA was used to prepare cDNA using iscript cDNA synthesis kit (1708891, Bio-rad). cDNA was diluted 1:5 before use.

SYBR Green Assay:

Article Title: The Ataxia telangiectasia-mutated and Rad3-related Protein Kinase Regulates Cellular Hydrogen Sulfide Concentrations
Article Snippet: One μg of RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-rad, 1708891). .. Quantitative real-time polymerase chain reaction was performed using SYBR Green Master Mix (Bio-rad, 1708882), and gene expression was quantified using the 2−ΔΔCT method.

Article Title: Endophytes alleviate the elevated CO2-dependent decrease in photosynthesis in rice, particularly under nitrogen limitation
Article Snippet: .. The isolated mRNAs were mixed with a master reaction mixture containing fluorescent dyes (iQ SyBR Green Supermix, Bio-Rad Laboratories) prior to cDNA synthesis using an iScript cDNA Synthesis Kit (Bio-Rad Laboratories) and were then loaded into a real-time qPCR thermocycler (Chromo 4 System, Bio-Rad Laboratories). .. The relative gene expression of rbcS was calculated according to .

Article Title: Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
Article Snippet: Five hours after removal of bacteria, RNA was extracted directly from monolayers within the 12-well plates using the RNeasy Micro Kit (QIAGEN, Venlo, The Netherlands) according to the manufacturer’s instructions and converted to cDNA using the iSCRIPT cDNA Synthesis Kit (BioRad, Hercules, CA, USA). .. Quantitative real-time PCR (qPCR) Master-mix was prepared as follows: 5 µL Iq Sybr green Master-mix (Biorad), 1 µL of forward and reverse primers each (final concentration 100 nM), 1 µL of demineralized water and 2 µL of 1:20 diluted cDNA.

Article Title: Inflammation and TGF-β Signaling Differ between Abdominal Aneurysms and Occlusive Disease
Article Snippet: .. Total RNA was reverse transcribed using iScript cDNA Synthesis Kit (Bio-Rad, Veenendaal, The Netherlands). cDNA samples were subjected to 40 cycles real-time PCR analysis using SYBR Green qPCR Master Mix 2x (Bio-Rad, Veenendaal, The Netherlands) and primers. ..

Article Title: Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating P38 Mitogen-Activated Protein Kinase Expression
Article Snippet: The cDNA was synthesized from 1 μg of RNA by applying to the Bio-Rad iScript cDNA synthesis kit (Bio-Rad). .. Quantitative polymerase chain reaction was performed on the MJ Opticon (Bio-Rad) system using iTaq Universal SYBR Green Supermix (Bio-Rad).

Article Title: Deficiency of T cell CD40L has minor beneficial effects on obesity-induced metabolic dysfunction
Article Snippet: All RNA was reverse transcribed to cDNA using an iScript cDNA synthesis kit (Bio-Rad, Lunteren, The Netherlands). .. Quantitative PCR was performed with a SYBR green PCR kit (Applied Biosystems, Waltham) on a ViiA7 real-time PCR system (Applied Biosystems).

Article Title: Long-chain acyl-CoA synthetase 1 interacts with key proteins that activate and direct fatty acids into niche hepatic pathways
Article Snippet: .. Total RNA and DNA were simultaneously isolated from liver using 0.7 ml of TRIzol (ThermoFisher Scientific) per 200 mg of tissue. cDNA was synthesized using 1 μg of RNA template (iScript cDNA Synthesis kit, Bio-Rad), and 50 ng of cDNA was added to each qPCR with SYBR Green (iTaq Universal SYBR; Bio-Rad) and used to detect amplicons with primers specific to the gene of interest (S6) and quantified using a qPCR thermocycler (Bio-Rad). ..

Article Title: Age-Related Changes in HAPLN1 Increase Lymphatic Permeability and Affect Routes of Melanoma Metastasis
Article Snippet: One microgram of RNA was used to prepare cDNA using iscript cDNA synthesis kit (1708891, Bio-rad). cDNA was diluted 1:5 before use. .. Each 20μl reaction comprised 1μl Power SYBR Green Master Mix (4367659, Invitrogen), 1μl primer mix (Final concentration 0.5μM) and 1μl cDNA.

Article Title: Improvement of epidermal covering on AEC patients with severe skin erosions by PRIMA-1MET/APR-246
Article Snippet: RNA was then extracted using RNEasy Mini kit (Qiagen), and cDNA was synthesized from 1 µg of RNA using iScript cDNA synthesis kit (Bio-Rad). .. Quantitative PCR was performed in triplicate using 2× SYBR Green PCR Master Mix (Absource Biotools).

Activation Assay:

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions. .. The results were normalized against the mean expression of two housekeeping genes: YWHAZ (tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta, Hs03044281_g1) and ACTB (β -actin, Hs99999903_m1) identified in a previous study [ ].

Generated:

Article Title: Age-Related Changes in HAPLN1 Increase Lymphatic Permeability and Affect Routes of Melanoma Metastasis
Article Snippet: One microgram of RNA was used to prepare cDNA using iscript cDNA synthesis kit (1708891, Bio-rad). cDNA was diluted 1:5 before use. .. Standard curves were generated for each primer and used to perform relative quantification.

Quantitative RT-PCR:

Article Title: Differentiation of diabetic foot ulcer–derived induced pluripotent stem cells reveals distinct cellular and tissue phenotypes
Article Snippet: Paragraph title: Quantitative RT-PCR ... RNA was reverse transcribed with the iScript cDNA Synthesis Kit (170-8891; Bio-Rad Laboratories, Hercules, CA, USA).

Article Title: MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages
Article Snippet: Paragraph title: Quantitative RT-PCR (Q-RT-PCR) ... Cells were incubated for the indicated times or overnight, after which time RNA was isolated using Trizol, then treated with DNAase1 (Invitrogen). cDNA was synthesized from 1 μg of RNA using iScript cDNA Synthesis Kit (BioRad).

Article Title: Improvement of epidermal covering on AEC patients with severe skin erosions by PRIMA-1MET/APR-246
Article Snippet: Paragraph title: qRT-PCR analyses ... RNA was then extracted using RNEasy Mini kit (Qiagen), and cDNA was synthesized from 1 µg of RNA using iScript cDNA synthesis kit (Bio-Rad).

Expressing:

Article Title: Restoring colistin sensitivity in colistin-resistant E. coli: Combinatorial use of MarR inhibitor with efflux pump inhibitor
Article Snippet: To discern the effect of expression efflux pump genes in colistin resistant U3790, we performed semi-quantitative gene expression studies by amplifying acrA, acrB and tolC genes in U3790. .. Total RNA from the bacterial strains were isolated using Aurum™ total RNA mini kit (Bio Rad). cDNA conversion was done using the iScript™ cDNA synthesis kit (Bio Rad).

Article Title: The Ataxia telangiectasia-mutated and Rad3-related Protein Kinase Regulates Cellular Hydrogen Sulfide Concentrations
Article Snippet: One μg of RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-rad, 1708891). .. Quantitative real-time polymerase chain reaction was performed using SYBR Green Master Mix (Bio-rad, 1708882), and gene expression was quantified using the 2−ΔΔCT method.

Article Title: Endophytes alleviate the elevated CO2-dependent decrease in photosynthesis in rice, particularly under nitrogen limitation
Article Snippet: Paragraph title: Gene expression analysis using dye-based qPCR ... The isolated mRNAs were mixed with a master reaction mixture containing fluorescent dyes (iQ SyBR Green Supermix, Bio-Rad Laboratories) prior to cDNA synthesis using an iScript cDNA Synthesis Kit (Bio-Rad Laboratories) and were then loaded into a real-time qPCR thermocycler (Chromo 4 System, Bio-Rad Laboratories).

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: Paragraph title: 2.3. RNA Isolation and Gene Expression ... Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions.

Article Title: Inflammation and TGF-β Signaling Differ between Abdominal Aneurysms and Occlusive Disease
Article Snippet: 2.1.7. qPCR Analysis Expression data of COL11A, Adiponectin, CXCL13, SLC7A5, and FDC-SP were analyzed in diseased aortic tissue ( ). .. Total RNA was reverse transcribed using iScript cDNA Synthesis Kit (Bio-Rad, Veenendaal, The Netherlands). cDNA samples were subjected to 40 cycles real-time PCR analysis using SYBR Green qPCR Master Mix 2x (Bio-Rad, Veenendaal, The Netherlands) and primers.

Article Title: Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating P38 Mitogen-Activated Protein Kinase Expression
Article Snippet: The cDNA was synthesized from 1 μg of RNA by applying to the Bio-Rad iScript cDNA synthesis kit (Bio-Rad). .. The relative expression of mRNA was calculated as: target gene = value of target gene / value of β-actin.

Article Title: Wnt-3a Induces Cytokine Release in Human Mast Cells
Article Snippet: .. Quantitative PCR (qPCR) and RNA Sequencing (RNAseq) For qPCR, RNA was extracted using an RNeasy Plus Mini Kit (Qiagen, Hilden, Germany), cDNA was prepared using an iScript cDNA synthesis kit (Bio-Rad), and qPCR was performed using TaqMan™ Gene Expression master mix (Thermo Fisher Scientific) on a CFX96 Real-Time System (Bio-Rad). .. The following primers were used: Hs02800695_m1 (HPRT1), Hs00174103_m1 (CXCL8), Hs04187715_m1 (CCL8), Hs00171147_m1 (CCL7), Hs01099660_g1 (CXCL5), Hs00234142_m1 (CCL3), and TaqMan Array Human WNT Pathway (all from Thermo Fisher Scientific).

Article Title: Deficiency of T cell CD40L has minor beneficial effects on obesity-induced metabolic dysfunction
Article Snippet: Paragraph title: Gene expression analysis ... All RNA was reverse transcribed to cDNA using an iScript cDNA synthesis kit (Bio-Rad, Lunteren, The Netherlands).

Article Title: Differentiation of diabetic foot ulcer–derived induced pluripotent stem cells reveals distinct cellular and tissue phenotypes
Article Snippet: RNA was reverse transcribed with the iScript cDNA Synthesis Kit (170-8891; Bio-Rad Laboratories, Hercules, CA, USA). .. Gene expression was quantified by the cycle threshold ΔΔ Ct method.

Article Title: Improvement of epidermal covering on AEC patients with severe skin erosions by PRIMA-1MET/APR-246
Article Snippet: RNA was then extracted using RNEasy Mini kit (Qiagen), and cDNA was synthesized from 1 µg of RNA using iScript cDNA synthesis kit (Bio-Rad). .. Expression of each gene was calculated using the 2−ΔΔCt method.

Polymerase Chain Reaction:

Article Title: Restoring colistin sensitivity in colistin-resistant E. coli: Combinatorial use of MarR inhibitor with efflux pump inhibitor
Article Snippet: Total RNA from the bacterial strains were isolated using Aurum™ total RNA mini kit (Bio Rad). cDNA conversion was done using the iScript™ cDNA synthesis kit (Bio Rad). .. The obtained PCR product was quantified using ImageJ software.

Article Title: The Ataxia telangiectasia-mutated and Rad3-related Protein Kinase Regulates Cellular Hydrogen Sulfide Concentrations
Article Snippet: One μg of RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-rad, 1708891). .. The primers used in polymerase chain reaction reactions are listed in .

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions. .. Real-time PCR was performed starting from 10 ng of cDNA, using a PCR mix containing TaqMan® Universal PCR Master Mix and Assays-on-Demand Gene expression probes (Life Technologies, Waltham, MA, USA) for SCX (scleraxis; Hs03054634_g1), COL1A1 (collagen type I alpha 1 chain; Hs01076777_m1), COL3A1 (collagen type III alpha 1 chain; Hs00943809_m1), MMP1 (metalloproteinase-1; Hs00899658_m1), MMP3 (metalloproteinase-3; Hs00968305_m1), and PTGS2 (cyclooxygenase-2; Hs00153133_m1).

Article Title: Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
Article Snippet: Five hours after removal of bacteria, RNA was extracted directly from monolayers within the 12-well plates using the RNeasy Micro Kit (QIAGEN, Venlo, The Netherlands) according to the manufacturer’s instructions and converted to cDNA using the iSCRIPT cDNA Synthesis Kit (BioRad, Hercules, CA, USA). .. For qPCR, a three-step PCR protocol was used: An initial 10 min denaturation at 95 °C, followed by 40 cycles with 15 s of denaturation at 95 °C, 30 s annealing at primer specific temperature, and 30 s of elongation at 72 °C.

Article Title: Differentiation of diabetic foot ulcer–derived induced pluripotent stem cells reveals distinct cellular and tissue phenotypes
Article Snippet: RNA was reverse transcribed with the iScript cDNA Synthesis Kit (170-8891; Bio-Rad Laboratories, Hercules, CA, USA). .. For each PCR reaction, 0.5 μl of cDNA was used.

Article Title: Improvement of epidermal covering on AEC patients with severe skin erosions by PRIMA-1MET/APR-246
Article Snippet: RNA was then extracted using RNEasy Mini kit (Qiagen), and cDNA was synthesized from 1 µg of RNA using iScript cDNA synthesis kit (Bio-Rad). .. Quantitative PCR was performed in triplicate using 2× SYBR Green PCR Master Mix (Absource Biotools).

Real-time Polymerase Chain Reaction:

Article Title: The Ataxia telangiectasia-mutated and Rad3-related Protein Kinase Regulates Cellular Hydrogen Sulfide Concentrations
Article Snippet: Paragraph title: 2.6. Quantitative Real-Time Polymerase Chain Reaction ... One μg of RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-rad, 1708891).

Article Title: Endophytes alleviate the elevated CO2-dependent decrease in photosynthesis in rice, particularly under nitrogen limitation
Article Snippet: .. The isolated mRNAs were mixed with a master reaction mixture containing fluorescent dyes (iQ SyBR Green Supermix, Bio-Rad Laboratories) prior to cDNA synthesis using an iScript cDNA Synthesis Kit (Bio-Rad Laboratories) and were then loaded into a real-time qPCR thermocycler (Chromo 4 System, Bio-Rad Laboratories). .. The relative gene expression of rbcS was calculated according to .

Article Title: Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an In Vitro Model of Tendon Cell Inflammation
Article Snippet: Total RNA from each sample was transcribed to cDNA using iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA, USA) as per manufacturer's instructions. .. Real-time PCR was performed starting from 10 ng of cDNA, using a PCR mix containing TaqMan® Universal PCR Master Mix and Assays-on-Demand Gene expression probes (Life Technologies, Waltham, MA, USA) for SCX (scleraxis; Hs03054634_g1), COL1A1 (collagen type I alpha 1 chain; Hs01076777_m1), COL3A1 (collagen type III alpha 1 chain; Hs00943809_m1), MMP1 (metalloproteinase-1; Hs00899658_m1), MMP3 (metalloproteinase-3; Hs00968305_m1), and PTGS2 (cyclooxygenase-2; Hs00153133_m1).

Article Title: Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
Article Snippet: Total RNA Extraction and Reverse Transcription Transcription levels of TNFα, SAA3, NF-κB and TLR-2 genes of PS in response to randomly selected CC151 (n = 4), CC479 (n = 4) and CC133 (n = 4) isolates were measured using quantitative real-time PCR (qPCR). .. Five hours after removal of bacteria, RNA was extracted directly from monolayers within the 12-well plates using the RNeasy Micro Kit (QIAGEN, Venlo, The Netherlands) according to the manufacturer’s instructions and converted to cDNA using the iSCRIPT cDNA Synthesis Kit (BioRad, Hercules, CA, USA).

Article Title: Inflammation and TGF-β Signaling Differ between Abdominal Aneurysms and Occlusive Disease
Article Snippet: .. Total RNA was reverse transcribed using iScript cDNA Synthesis Kit (Bio-Rad, Veenendaal, The Netherlands). cDNA samples were subjected to 40 cycles real-time PCR analysis using SYBR Green qPCR Master Mix 2x (Bio-Rad, Veenendaal, The Netherlands) and primers. ..

Article Title: Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating P38 Mitogen-Activated Protein Kinase Expression
Article Snippet: Paragraph title: Quantitative Real-Time Polymerase Chain Reaction ... The cDNA was synthesized from 1 μg of RNA by applying to the Bio-Rad iScript cDNA synthesis kit (Bio-Rad).

Article Title: Wnt-3a Induces Cytokine Release in Human Mast Cells
Article Snippet: .. Quantitative PCR (qPCR) and RNA Sequencing (RNAseq) For qPCR, RNA was extracted using an RNeasy Plus Mini Kit (Qiagen, Hilden, Germany), cDNA was prepared using an iScript cDNA synthesis kit (Bio-Rad), and qPCR was performed using TaqMan™ Gene Expression master mix (Thermo Fisher Scientific) on a CFX96 Real-Time System (Bio-Rad). .. The following primers were used: Hs02800695_m1 (HPRT1), Hs00174103_m1 (CXCL8), Hs04187715_m1 (CCL8), Hs00171147_m1 (CCL7), Hs01099660_g1 (CXCL5), Hs00234142_m1 (CCL3), and TaqMan Array Human WNT Pathway (all from Thermo Fisher Scientific).

Article Title: Deficiency of T cell CD40L has minor beneficial effects on obesity-induced metabolic dysfunction
Article Snippet: All RNA was reverse transcribed to cDNA using an iScript cDNA synthesis kit (Bio-Rad, Lunteren, The Netherlands). .. Quantitative PCR was performed with a SYBR green PCR kit (Applied Biosystems, Waltham) on a ViiA7 real-time PCR system (Applied Biosystems).

Article Title: Long-chain acyl-CoA synthetase 1 interacts with key proteins that activate and direct fatty acids into niche hepatic pathways
Article Snippet: .. Total RNA and DNA were simultaneously isolated from liver using 0.7 ml of TRIzol (ThermoFisher Scientific) per 200 mg of tissue. cDNA was synthesized using 1 μg of RNA template (iScript cDNA Synthesis kit, Bio-Rad), and 50 ng of cDNA was added to each qPCR with SYBR Green (iTaq Universal SYBR; Bio-Rad) and used to detect amplicons with primers specific to the gene of interest (S6) and quantified using a qPCR thermocycler (Bio-Rad). ..

Article Title: Age-Related Changes in HAPLN1 Increase Lymphatic Permeability and Affect Routes of Melanoma Metastasis
Article Snippet: Paragraph title: Quantitative PCR ... One microgram of RNA was used to prepare cDNA using iscript cDNA synthesis kit (1708891, Bio-rad). cDNA was diluted 1:5 before use.

Article Title: Improvement of epidermal covering on AEC patients with severe skin erosions by PRIMA-1MET/APR-246
Article Snippet: RNA was then extracted using RNEasy Mini kit (Qiagen), and cDNA was synthesized from 1 µg of RNA using iScript cDNA synthesis kit (Bio-Rad). .. Quantitative PCR was performed in triplicate using 2× SYBR Green PCR Master Mix (Absource Biotools).

Recombinant:

Article Title: MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages
Article Snippet: Treatment groups included DMEM Complete alone or DMEM Complete with 100ng/ml recombinant murine Mst1 (R & D 6244-MS). .. Cells were incubated for the indicated times or overnight, after which time RNA was isolated using Trizol, then treated with DNAase1 (Invitrogen). cDNA was synthesized from 1 μg of RNA using iScript cDNA Synthesis Kit (BioRad).

Software:

Article Title: Restoring colistin sensitivity in colistin-resistant E. coli: Combinatorial use of MarR inhibitor with efflux pump inhibitor
Article Snippet: Total RNA from the bacterial strains were isolated using Aurum™ total RNA mini kit (Bio Rad). cDNA conversion was done using the iScript™ cDNA synthesis kit (Bio Rad). .. The obtained PCR product was quantified using ImageJ software.

Article Title: Hypoxia-Induced Phenotypic Transformation of Corpus Cavernosum Smooth Muscle Cells After Cavernous Nerve Crush Injury by Down-Regulating P38 Mitogen-Activated Protein Kinase Expression
Article Snippet: The rat α-SMA, vimentin, ERK, JNK, p38, and β-actin sequences reported in the National Center for Biotechnology Information gene database were designed using Oligo 6.71 primer analysis software, and then were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). .. The cDNA was synthesized from 1 μg of RNA by applying to the Bio-Rad iScript cDNA synthesis kit (Bio-Rad).

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    Bio-Rad iscript cdna synthesis kit
    RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to <t>cDNA</t> using <t>iScript</t> cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.
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    RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to cDNA using iScript cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.

    Journal: Bio-protocol

    Article Title: Immunoprecipitation of Tri-methylated Capped RNA

    doi: 10.21769/BioProtoc.2717

    Figure Lengend Snippet: RNA immunoprecipitation of (TMG)-capped primary miRNAs (Pri-miRNAs) in quiescent human foreskin fibroblasts (HFFs) RT-PCR data shows the RNA immunoprecipitation of Pri-miR-34a and Pri-miR-3188 in quiescent HFFs (as well as the positive control snRNA U7), but not Pri-miR-423 using an antibody against (TMG)-capped RNAs. Total RNA was extracted using TRIzol Reagent, and 10 μg of RNA was diluted in NET-2 buffer, precleared and incubated with Protein G Sepharose 4 Fast Flow beads loaded with 15 μl of control antibody (Normal Rabbit Serum, EMD-Millipore) or with antibody recognizing the (TMG)-cap (Anti-m3G-cap, rabbit polyclonal, Synaptic Systems). Beads were rinsed five times with NET-2 buffer and were resuspended in G-50 buffer. RNA was extracted from the beads by phenol-chloroform-isoamyl alcohol extraction and resuspended in 20 μl of nuclease-free water. Immunoprecipitated tri-methylated capped RNA was converted to cDNA using iScript cDNA synthesis kit (Bio-Rad), followed by RT-PCR, and visualized after gel electrophoresis.

    Article Snippet: Protein G Sepharose® 4 Fast Flow Beads (GE Healthcare, catalog number: 17061801) Normal rabbit serum (control, EMD Millipore, catalog number: NS01L-1ML) Sodium chloride (NaCl) (Fisher Scientific, catalog number: S671-3) NP-40 (Thermo Fisher Scientific, catalog number: 28324) Tris base (Fisher Scientific, catalog number: BP152-5) Hydrochloric acid (HCl) (VWR, catalog number: BDH7204-1) RNasin™ Plus RNase inhibitor (Promega, catalog number: N2611) NaOAc (AMRESCO, catalog number: 0602) Ethylenediaminetetraacetate acid (EDTA), pH 8 (Thermo Fisher Scientific, catalog number: AM9260G) Ethylenediaminetetraacetate acid (EDTA) (AMRESCO, catalog number: 0105) Sodium dodecyl sulfate (SDS) (Thermo Fisher Scientific, catalog number: AM9822) Phenol/Chloroform/Isoamyl Alcohol; 125:24:1 mixture, pH 4.5 (Thermo Fisher Scientific, catalog number: AM9720) Agarose LE (Denville Scientific, catalog number: CA3510-8) iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, catalog number: 1708891) Sso Advanced™ Universal SYBR® Green Supermix (Bio-Rad Laboratories, catalog number: 1725274) PARIS™ Kit (Thermo Fisher Scientific, catalog number: AM1921) Boric acid (Fisher Scientific, catalog number: A73-1) NET-2 Buffer (see Recipes) G-50 Buffer (see Recipes) 1× TBE (see Recipes)

    Techniques: Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Positive Control, Incubation, Flow Cytometry, Methylation, Nucleic Acid Electrophoresis

    Induction of autophagic genes expression following 4 weeks of nitrite therapy in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of autophagic markers beclin‐1 (A), ATG ‐5 (B), and ATG ‐7 (C). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ATG indicates autophagy related gene; CHF, chronic heart failure.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Nitrite Therapy Ameliorates Myocardial Dysfunction via H2S and Nuclear Factor‐Erythroid 2‐Related Factor 2 (Nrf2)‐Dependent Signaling in Chronic Heart Failure

    doi: 10.1161/JAHA.116.003551

    Figure Lengend Snippet: Induction of autophagic genes expression following 4 weeks of nitrite therapy in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of autophagic markers beclin‐1 (A), ATG ‐5 (B), and ATG ‐7 (C). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ATG indicates autophagy related gene; CHF, chronic heart failure.

    Article Snippet: One microgram of RNA was transcribed using an I‐script cDNA synthesis kit from Bio‐Rad.

    Techniques: Expressing, Mouse Assay, Software

    Effects of nitrite therapy on hypertrophic gene expression in ischemia‐induced CHF mice. Nitrite (100 mg/L) was given in the drinking water during reperfusion. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of ANP (A), BNP (B), and Myh7 (C) using TaqMan PCR assay system. The number inside the bar denotes the number of animals used per experiment. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ANP indicates atrial natriuretic peptide; BNP, brain natriuretic peptide; Myh7, myosin heavy chain beta; PCR, polymerase chain reaction.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Nitrite Therapy Ameliorates Myocardial Dysfunction via H2S and Nuclear Factor‐Erythroid 2‐Related Factor 2 (Nrf2)‐Dependent Signaling in Chronic Heart Failure

    doi: 10.1161/JAHA.116.003551

    Figure Lengend Snippet: Effects of nitrite therapy on hypertrophic gene expression in ischemia‐induced CHF mice. Nitrite (100 mg/L) was given in the drinking water during reperfusion. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of ANP (A), BNP (B), and Myh7 (C) using TaqMan PCR assay system. The number inside the bar denotes the number of animals used per experiment. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). ANP indicates atrial natriuretic peptide; BNP, brain natriuretic peptide; Myh7, myosin heavy chain beta; PCR, polymerase chain reaction.

    Article Snippet: One microgram of RNA was transcribed using an I‐script cDNA synthesis kit from Bio‐Rad.

    Techniques: Expressing, Mouse Assay, Aqueous Normal-phase Chromatography, Polymerase Chain Reaction, Software

    Effects of nitrite on cardiac antioxidant gene and protein levels in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of SOD 1 (A), catalase (B), and GPX (C) using TaqMan PCR assay system. (D) represents proteins levels of SOD 1, catalase, and GPX , and (E), (F), and (G) represent the quantitation of the blots in (D). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). CHF indicates chronic heart failure; GPX, glutathione peroxidase; PCR, polymerase chain reaction; SOD1, superoxide dismutase 1.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Nitrite Therapy Ameliorates Myocardial Dysfunction via H2S and Nuclear Factor‐Erythroid 2‐Related Factor 2 (Nrf2)‐Dependent Signaling in Chronic Heart Failure

    doi: 10.1161/JAHA.116.003551

    Figure Lengend Snippet: Effects of nitrite on cardiac antioxidant gene and protein levels in CHF mice. cDNA was prepared from RNA obtained from mouse heart tissues followed by analysis of mRNA of SOD 1 (A), catalase (B), and GPX (C) using TaqMan PCR assay system. (D) represents proteins levels of SOD 1, catalase, and GPX , and (E), (F), and (G) represent the quantitation of the blots in (D). The number in the circle inside the bar denotes the number of animals used. Differences in data between the groups were compared using Prism 6 (GraphPad Software, La Jolla, CA) with nonparametric test (Wilcoxon rank sum test). CHF indicates chronic heart failure; GPX, glutathione peroxidase; PCR, polymerase chain reaction; SOD1, superoxide dismutase 1.

    Article Snippet: One microgram of RNA was transcribed using an I‐script cDNA synthesis kit from Bio‐Rad.

    Techniques: Mouse Assay, Polymerase Chain Reaction, Quantitation Assay, Software

    Effect of silencing Schistosoma mansoni CYP450 in cultured larval worms. Freshly prepared schistosomula (300–400) were placed in each well containing 1 ml Basch’s Media in a 24-well plate and overnight in a 37°C with 5% CO2. The following day schistosomula were treated with 10 or 30 μg/ml S . mansoni CYP450 dsRNA or 30 μg/ml negative control dsRNA. Over several days worms were observed for dead (dark, granular appearance) or alive (translucent). (A) mRNA expression patterns in schistosomula treated with S . mansoni CYP450 specific dsRNA or negative control dsRNA control after 3 days of treatment (Experiments 1 and 2) or 2 days treatment (Experiment 3). The control gene for cDNA input is S . mansoni glyceraldehyde 3-phosphate dehydrogenase (GAPDH). C, schistosomula treated with 30 μg/mL irrelevant dsRNA; 10, schistosomula treated with 10 μg/mL Sm CYP450 dsRNA; 30, schistosomula treated with 30 μg/mL Sm CYP450 dsRNA. (B) Effect of S . mansoni CYP450 dsRNA on schistosomula survival in cultures with 30 μg/mL negative control dsRNA (black square), 10 μg/mL S . mansoni CYP450-specific ds RNA (open triangle), and 30 μg/mL S . mansoni CYP450-specific ds RNA (black triangle). Treatments were done in triplicate and repeated 3 times. Error bars indicate standard error of the mean; *, p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: The Schistosoma mansoni Cytochrome P450 (CYP3050A1) Is Essential for Worm Survival and Egg Development

    doi: 10.1371/journal.pntd.0004279

    Figure Lengend Snippet: Effect of silencing Schistosoma mansoni CYP450 in cultured larval worms. Freshly prepared schistosomula (300–400) were placed in each well containing 1 ml Basch’s Media in a 24-well plate and overnight in a 37°C with 5% CO2. The following day schistosomula were treated with 10 or 30 μg/ml S . mansoni CYP450 dsRNA or 30 μg/ml negative control dsRNA. Over several days worms were observed for dead (dark, granular appearance) or alive (translucent). (A) mRNA expression patterns in schistosomula treated with S . mansoni CYP450 specific dsRNA or negative control dsRNA control after 3 days of treatment (Experiments 1 and 2) or 2 days treatment (Experiment 3). The control gene for cDNA input is S . mansoni glyceraldehyde 3-phosphate dehydrogenase (GAPDH). C, schistosomula treated with 30 μg/mL irrelevant dsRNA; 10, schistosomula treated with 10 μg/mL Sm CYP450 dsRNA; 30, schistosomula treated with 30 μg/mL Sm CYP450 dsRNA. (B) Effect of S . mansoni CYP450 dsRNA on schistosomula survival in cultures with 30 μg/mL negative control dsRNA (black square), 10 μg/mL S . mansoni CYP450-specific ds RNA (open triangle), and 30 μg/mL S . mansoni CYP450-specific ds RNA (black triangle). Treatments were done in triplicate and repeated 3 times. Error bars indicate standard error of the mean; *, p

    Article Snippet: Total RNA was used for cDNA synthesis (iScript, BIO-RAD) per the manufacturer’s recommendation.

    Techniques: Cell Culture, Negative Control, Expressing

    CYP450 messenger RNA abundance during the lifecycle of Schistosoma mansoni . Whole RNA was extracted from different stages of S . mansoni (cercariae, 1-day old schistosomula; juvenile liver worms (23 days post infection), adult males (49 days post infection), adult females (49 days post infection) and eggs) using TRIzol reagent and chloroform/ethanol extraction protocol. cDNA was synthesized from whole RNA and used for qRT-PCR, with reactions done in triplicate. Adult males (= 1) were used as calibrator stage and mRNA abundance was normalized to α-tubulin. Error bars indicate standard error of the mean with n ≥ 3 biological replicates. Numbers indicate fold change relative to adult males and all values are significantly different from adult males; p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: The Schistosoma mansoni Cytochrome P450 (CYP3050A1) Is Essential for Worm Survival and Egg Development

    doi: 10.1371/journal.pntd.0004279

    Figure Lengend Snippet: CYP450 messenger RNA abundance during the lifecycle of Schistosoma mansoni . Whole RNA was extracted from different stages of S . mansoni (cercariae, 1-day old schistosomula; juvenile liver worms (23 days post infection), adult males (49 days post infection), adult females (49 days post infection) and eggs) using TRIzol reagent and chloroform/ethanol extraction protocol. cDNA was synthesized from whole RNA and used for qRT-PCR, with reactions done in triplicate. Adult males (= 1) were used as calibrator stage and mRNA abundance was normalized to α-tubulin. Error bars indicate standard error of the mean with n ≥ 3 biological replicates. Numbers indicate fold change relative to adult males and all values are significantly different from adult males; p

    Article Snippet: Total RNA was used for cDNA synthesis (iScript, BIO-RAD) per the manufacturer’s recommendation.

    Techniques: Infection, Synthesized, Quantitative RT-PCR