Revertaid first strand cdna synthesis reagents

Manufactured by Thermo Fisher Scientific

Sourced in United States

RevertAid First Strand cDNA Synthesis reagents are a set of reagents used for the reverse transcription of RNA into complementary DNA (cDNA). The reagents include a reverse transcriptase enzyme, reaction buffer, and necessary components for the cDNA synthesis process.

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Lab products found in correlation

Maxima sybr green master mix, by Thermo Fisher Scientific (2 mentions) Total rna zol out kit, by A&A Biotechnology (1 mentions) Taqman microrna assay, by Thermo Fisher Scientific (1 mentions) Hot firepol evagreen qpcr supermix, by Solis BioDyne (1 mentions) Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Direct zol rna miniprep kit, by Zymo Research (1 mentions) Mirneasy mini kit, by Qiagen (1 mentions) Abi prism 7900ht system, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)

3 protocols using revertaid first strand cdna synthesis reagents

RNA Extraction and RT-qPCR Analysis

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Total RNA was extracted with Direct-zol™ RNA MiniPrep Kit (Zymo
Research, Irvine, CA, USA), miRNeasy Mini Kit (Qiagen, Hilden, Germany) or Total
RNA Zol-Out Kit (A&A Biotechnology, Gdynia, Poland). Reverse transcription
was performed with RevertAid First Strand cDNA Synthesis reagents (Thermo Fisher
Scientific, Waltham, MA, USA) for mRNA analysis or TaqMan® MicroRNA
Reverse Transcription Kit and TaqMan™ MicroRNA Assays (both from Thermo
Fisher Scientific) for microRNA analysis. RT-qPCR was done using custom primers
and 5x HOT FIREPol EvaGreen qPCR Supermix (Solis BioDyne, Tartu, Estonia) for
mRNA analysis or TaqMan™ MicroRNA Assays and 5 x HOT FIREPol®
Probe qPCR Mix Plus (ROX) (Solis BioDyne) for miRNA analysis. Primer sequences
and additional information are provided in Supporting Methods.

Vaher H., Runnel T., Urgard E., Aab A., Carreras Badosa G., Maslovskaja J., Abram K., Raam L., Kaldvee B., Annilo T., Tkaczyk E.R., Maimets T., Akdis C.A., Kingo K, & Rebane A. (2019). miR-10a-5p is increased in atopic dermatitis and has capacity to inhibit keratinocyte proliferation. Allergy, 74(11), 2146-2156.

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Reverse Transcription and qPCR for Tissue RNA Analysis

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For mouse organ tissue, cells and human patient tissue, RNA was reverse-transcribed using the RevertAid First Strand cDNA Synthesis reagents (Thermo Fisher Scientific), and RT–qPCR was performed in duplicates or triplicates using the Maxima SYBR Green Master Mix (Thermo Fisher Scientific) on QuantStudio 6/7 qPCR instruments. A list of the primer sequences used is provided in Supplementary Table 1.

Gulen M.F., Samson N., Keller A., Schwabenland M., Liu C., Glück S., Thacker V.V., Favre L., Mangeat B., Kroese L.J., Krimpenfort P., Prinz M, & Ablasser A. (2023). cGAS–STING drives ageing-related inflammation and neurodegeneration. Nature, 620(7973), 374-380.

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Quantitative PCR for Mouse Lung Samples

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For mouse lung samples, RNA was reverse-transcribed using the RevertAid First Strand cDNA Synthesis reagents (Thermo Fisher Scientific), and quantitative PCR with reverse transcription (RT–qPCR) was performed in duplicate using Maxima SYBR Green Master Mix (Thermo Fisher Scientific) on QuantStudio 6/7 qPCR instruments. For LoC samples, RNA was reverse-transcribed using the SuperScript IV First-Strand Synthesis System with random hexamers (Invitrogen), and RT–qPCR reactions were prepared with SYBR Green PCR Master Mix (Applied Biosystems) on the ABI PRISM 7900HT System (Applied Biosystems). Amplicon specificity was confirmed by melting-curve analysis. The primer sequences are listed in Supplementary Table 2.

Domizio J.D., Gulen M.F., Saidoune F., Thacker V.V., Yatim A., Sharma K., Nass T., Guenova E., Schaller M., Conrad C., Goepfert C., de Leval L., Garnier C.V., Berezowska S., Dubois A., Gilliet M, & Ablasser A. (2022). The cGAS–STING pathway drives type I IFN immunopathology in COVID-19. Nature, 603(7899), 145-151.

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