Quantification of mRNA Expression via RT-PCR

mRNA expression was quantitated using RT-PCR as previously described [5 (link),46 (link),47 (link),48 (link)]. Briefly, TRIzol (Life Technologies) extracted total RNA was quantified using a NanoDrop 1000 spectrophotometer (Thermo Scientific). Total RNA (250 ng) was subjected to cDNA synthesis using the High Efficiency cDNA Synthesis Kit (Life Technologies). The levels of various mRNA transcripts were determined using Fast SYBR Green (Life Technologies) and gene-specific forward and reverse primer sets (Table 1) on a ViiA7 instrument (Life Technologies). 18s RNA expression was used to normalize mRNA expression using the ΔΔCT comparative method.

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Hamid T., Ismahil M.A., Bansal S.S., Patel B., Goel M., White C.R., Anantharamaiah G.M, & Prabhu S.D. (2020). The Apolipoprotein A-I Mimetic L-4F Attenuates Monocyte Activation and Adverse Cardiac Remodeling after Myocardial Infarction. International Journal of Molecular Sciences, 21(10), 3519.

Publication 2020

TRIzol NanoDrop 1000 spectrophotometer High Efficiency cDNA Synthesis Kit Fast SYBR Green ViiA7 instrument

Cdna Fast green Gene Mrna Primer Rna expression Rt pcr Synthesis Trizol

Corresponding Organization : Birmingham VA Medical Center

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Variable analysis

independent variables

MRNA expression

dependent variables

Levels of various mRNA transcripts

control variables

18s RNA expression used to normalize mRNA expression

controls

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

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