Quantifying Gene Expression in Plants

Total RNA was extracted from 5 g samples of roots, stems, leaves and seeds, respectively, with the TaKaRa MiniBEST Plant RNA Extraction Kit (TaKaRa). First-strand cDNA was synthesized and qPCR was carried out using a LightCycler^®480 System Real-Time PCR as described by Feng et al. (2019) (link). All primers for the target genes are listed in Table 1. Relative expression of target genes was evaluated by the 2^–ΔΔCt method (Li et al., 2018 (link)). The experiments were conducted in three biological replications.

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Feng Y., Han Y., Han B., Zhao Y., Yang Y, & Xing Y. (2022). A 4 bp InDel in the Promoter of Wheat Gene TaAFP-B Affecting Seed Dormancy Confirmed in Transgenic Rice. Frontiers in Plant Science, 13, 837805.

Publication 2022

TaKaRa MiniBEST Plant RNA Extraction Kit

Biological Cdna Expression genes Genes Plant rna Primers Real time pcr Replications Roots Seeds Stems

Corresponding Organization : Inner Mongolia Agricultural University

Other organizations : Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Ministry of Agriculture and Rural Affairs, Henan Academy of Agricultural Sciences

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Variable analysis

independent variables

Tissue type (roots, stems, leaves, seeds)

dependent variables

Relative expression of target genes

control variables

Amount of input total RNA (5 g samples)
RNA extraction method (TaKaRa MiniBEST Plant RNA Extraction Kit)
CDNA synthesis method
Real-time PCR platform (LightCycler^®480 System)
PCR primers for target genes (listed in Table 1)
Data analysis method (2^–ΔΔCt method)

controls

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

Annotations

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