Quantification of Pepper Gene Expression

Total RNA extraction was conducted with the total RNA Miniprep Kit (Axygen Biosciences, Union City, CA, United States). Approximately 5 μg of RNA was reverse transcribed with the ReverTra Ace qPCR-RT Kit (Toyobo, Japan) according to the manufacturer’s protocol. qRT-PCR was carried out in triplicate using the iCycler iQ^TM Real-time PCR Detection System (Bio-Rad, Hercules, CA, United States) with the procedures as previously described (Yang et al., 2019 (link)). The pepper actin gene was used as an internal control. The primers of actin and CaPIF genes are listed in Supplementary Table 1.

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Yang Y., Guang Y., Wang F., Chen Y., Yang W., Xiao X., Luo S, & Zhou Y. (2021). Characterization of Phytochrome-Interacting Factor Genes in Pepper and Functional Analysis of CaPIF8 in Cold and Salt Stress. Frontiers in Plant Science, 12, 746517.

Publication 2021

Miniprep Kit ReverTra Ace qPCR-RT Kit iCycler iQTM Real-time PCR Detection System

Actin Genes Pepper Primers

Corresponding Organization : Jiangxi Agricultural University

Other organizations : Shenyang Agricultural University

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Variable analysis

independent variables

Total RNA extraction method (using total RNA Miniprep Kit)
Reverse transcription method (using ReverTra Ace qPCR-RT Kit)

dependent variables

Gene expression levels of target genes (CaPIF) measured by qRT-PCR

control variables

Pepper actin gene as an internal control

controls

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

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