RNA Extraction and cDNA Synthesis in Mouse Embryos

Total RNA was isolated from the yolk sacs and livers of mouse embryos using Trizol extraction (Life Technologies, Grand Island, NY, USA) according to manufacturer’s instructions, as previously described^{51 (link)}. Complementary DNA (cDNA) was prepared from 2 µg RNA using Maloney murine leukemia virus reverse transcriptase (Life Technologies) and an oligo(dT) primer according to the manufacturer's instructions. PCR was performed on 2 µl cDNA using Taq DNA polymerase (Promega, Madison, WI, USA) in an Eppendorf thermocycler^{51 (link)}.

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Belyea B.C., Santiago A.E., Vasconez W.A., Nagalakshmi V.K., Xu F., Mehalic T.C., Sequeira-Lopez M.L, & Gomez R.A. (2021). A primitive type of renin-expressing lymphocyte protects the organism against infections. Scientific Reports, 11, 7251.

Publication 2021

Trizol extraction Maloney murine leukemia virus reverse transcriptase Taq DNA polymerase thermocycler

Cdna Embryos Livers Mouse Murine leukemia virus Oligo Primer Promega Reverse transcriptase Taq dna polymerase Trizol Yolk sacs

Corresponding Organization :

Other organizations : University of Virginia

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Variable analysis

independent variables

Trizol extraction method
Maloney murine leukemia virus reverse transcriptase
Oligo(dT) primer
Taq DNA polymerase

dependent variables

Total RNA isolated from yolk sacs and livers of mouse embryos
Complementary DNA (cDNA) prepared from 2 µg RNA

control variables

Manufacturer's instructions for Trizol extraction, reverse transcription, and PCR
Previously described procedures^{51 (link)}

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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