Quantitative Real-time PCR Analysis

As previously described [46 (link)], total RNA was extracted with RNAiso Plus solution (TAKARA, Dalian, China, 9109), and cDNAs were synthesized using a PrimeScrip RT reagent Kit (TAKARA, RR037A). Quantitative Real-time PCR was performed on Thermo Scientific QuantStudio 3 Real-Time PCR System using FastStart Universal SYBR Green Master (Roche Life Science, Mannheim, Germany, 04913914001). The data were analyzed using the comparative threshold cycle (ΔΔCt) method and normalized to Gapdh. All PCR primers used are listed in Supplementary Table S2.

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Wang S., Qiao H., Wang P., Wang Y, & Qin D. (2021). ZDHHC19 Is Dispensable for Spermatogenesis, but Is Essential for Sperm Functions in Mice. International Journal of Molecular Sciences, 22(16), 8894.

Publication 2021

RNAiso Plus solution PrimeScrip RT reagent Kit QuantStudio 3 Real-Time PCR System FastStart Universal SYBR Green Master

Cdnas Gapdh Primers Quantitative real time pcr Sybr green

Corresponding Organization : Michigan State University

Other organizations : East China Normal University

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Gene expression levels

control variables

RNAiso Plus solution
PrimeScrip RT reagent Kit
FastStart Universal SYBR Green Master
Gapdh (for normalization)

positive controls

Not specified

negative controls

Not specified

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