Journal: Cell reports

Article Title: LKB1 and AMPK instruct cone nuclear position to modify visual function

doi: 10.1016/j.celrep.2021.108698

Figure Lengend Snippet:

Article Snippet: iScript Reverse Transcription Supermix for RT-qPCR , Bio-Rad , Cat# 1708840.

Techniques: Recombinant, SYBR Green Assay, Software

Journal: PLoS ONE

Article Title: An overview of technical considerations when using quantitative real-time PCR analysis of gene expression in human exercise research

doi: 10.1371/journal.pone.0196438

Figure Lengend Snippet: A. Summary of the key steps leading from gene to protein expression in eukaryotes. DNA is first transcribed into RNA, then processed to mRNA after removing the noncoding regions (introns, green) and splicing the coding regions (exons, red) together. The spliced mRNA (red) is then exported to the cytoplasm to produce the protein molecule. B. Outline of procedures for first strand complementary DNA (cDNA) synthesis from messenger RNA (mRNA) using short sequences of deoxy-thymidine nucleotides (oligo-dT primers, green). After annealing of Oligo-dT primers to the mRNA sequence (grey), the RNA-directed DNA polymerase, reverse transcriptase, is able to synthesise cDNA strand (blue), as well as degrade mRNA (red) from the hybrid molecule.

Article Snippet: For all of our experiments, we used Bio-Rad iScriptTM RT Supermix cDNA synthesis kit (Bio-Rad, Hercules, CA) to generate cDNA.

Techniques: Expressing, cDNA Synthesis, Sequencing, Reverse Transcription

Journal: PLoS ONE

Article Title: An overview of technical considerations when using quantitative real-time PCR analysis of gene expression in human exercise research

doi: 10.1371/journal.pone.0196438

Figure Lengend Snippet: Skeletal muscle samples are cleaned quickly and snap-frozen in liquid nitrogen after the muscle biopsy, and then stored at -80°C until RNA extraction. RNA is extracted using TRIzol or another Tri-reagent, and assessed for concentration and quality. RNA that passes the quality test is used to synthesise cDNA, which is used as template in the final qPCR assay. The conditions of the final qPCR assay must be optimised for each experiment, and the data should be processed using the correct analysis methods.

Article Snippet: For all of our experiments, we used Bio-Rad iScriptTM RT Supermix cDNA synthesis kit (Bio-Rad, Hercules, CA) to generate cDNA.

Techniques: RNA Extraction, Concentration Assay

Journal: PLoS ONE

Article Title: An overview of technical considerations when using quantitative real-time PCR analysis of gene expression in human exercise research

doi: 10.1371/journal.pone.0196438

Figure Lengend Snippet: DNA fragment products produced from a PCR reaction with the same primers, but using either water (Lane 2), -RT (cDNA synthesis reaction containing RNA but no cDNA, Lane 3), or cDNA (Lane 4) as template, were separated on a 2% agarose gel. Lane 1 is the 100 bp DNA ladder. A single sharp DNA band of the expected size, which is the final PCR amplicons, is present only in the reaction with cDNA.

Article Snippet: For all of our experiments, we used Bio-Rad iScriptTM RT Supermix cDNA synthesis kit (Bio-Rad, Hercules, CA) to generate cDNA.

Techniques: Produced, cDNA Synthesis, Agarose Gel Electrophoresis

Journal: PLoS ONE

Article Title: An overview of technical considerations when using quantitative real-time PCR analysis of gene expression in human exercise research

doi: 10.1371/journal.pone.0196438

Figure Lengend Snippet: The qPCR reaction using cDNA synthesised from Intact (pink) and Degraded RNA (red) sample show the same melting curve, indicating that the same PCR amplicon is produced. However, a different melting curve is observed when using cDNA synthesised from RNase Treated RNA sample (blue), which shows a different PCR amplicon is produced during the qPCR reaction. Our recommendations for qPCR optimisation are listed in .

Article Snippet: For all of our experiments, we used Bio-Rad iScriptTM RT Supermix cDNA synthesis kit (Bio-Rad, Hercules, CA) to generate cDNA.

Techniques: Amplification, Produced

Journal: PLoS ONE

Article Title: An overview of technical considerations when using quantitative real-time PCR analysis of gene expression in human exercise research

doi: 10.1371/journal.pone.0196438

Figure Lengend Snippet: A standard curve was generated using a 10-fold dilution of cDNA as template for qPCR reactions. The resulting C q values are plotted against the Log of the cDNA input. The efficiency, as well as the R 2 value, are within the acceptable range. The efficiencies of Cyclophilin and PGC-1α are approximately equal, as the absolute value of the slope of ΔC q against the Log of the cDNA input is < 0.1.

Article Snippet: For all of our experiments, we used Bio-Rad iScriptTM RT Supermix cDNA synthesis kit (Bio-Rad, Hercules, CA) to generate cDNA.

Techniques: Generated

Journal: PLoS ONE

Article Title: An overview of technical considerations when using quantitative real-time PCR analysis of gene expression in human exercise research

doi: 10.1371/journal.pone.0196438

Figure Lengend Snippet: A: Determination of cDNA amount in reactions with different RNA input. Different amounts of RNA were used to synthesise cDNA (n = 4 for each RNA input), and the relative amount of cDNA in each reaction was measured using OliGreen dye. Values are presented as mean ± SD. B: Correlation between RNA input and average relative amount of cDNA measured.

Article Snippet: For all of our experiments, we used Bio-Rad iScriptTM RT Supermix cDNA synthesis kit (Bio-Rad, Hercules, CA) to generate cDNA.

Techniques: