Comprehensive Transcriptome Analysis Pipeline

RNA extraction and reverse transcription were performed according to the instructions in the Plant RNA maxi kit (Biomiga, San Diego, CA, USA) and the EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMix (TransGen Biotech Co., Beijing, China), respectively. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) analysis was performed using TransStart Probe qPCR SuperMix (TransGen Biotech Co.) and detected using a CFX96 Touch™ Real-Time PCR Detection System (Bio-rad Laboratories, Inc., Singapore, Singapore). Relative expression levels were calculated using the 2^−△△Ct method [50 (link)]. All RT-qPCR primers are listed in Table S1. In this study, Tubulin (accession No. AB239680.1) was used as an internal reference [4 (link)]. The transcriptome sequencing data have been deposited in the Sequence Read Archive (https://www.ncbi.nlm.nih.gov/sra) under accession numbers SUB2967341 [6 (link)].

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Cheng X., Li G., Manzoor M.A., Wang H., Abdullah M., Su X., Zhang J., Jiang T., Jin Q., Cai Y, & Lin Y. (2019). In Silico Genome-Wide Analysis of Respiratory Burst Oxidase Homolog (RBOH) Family Genes in Five Fruit-Producing Trees, and Potential Functional Analysis on Lignification of Stone Cells in Chinese White Pear. Cells, 8(6), 520.

Publication 2019

EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMix TransStart Probe qPCR SuperMix CFX96 Touch™ Real-Time PCR Detection System

Cdna Plant rna Primers Quantitative real time polymerase chain reaction Reverse transcription Synthesis Touch Tubulin

Corresponding Organization : Anhui Agricultural University

Other organizations : Anhui Academy of Agricultural Sciences

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Relative expression levels of target genes measured by RT-qPCR

control variables

RNA extraction and reverse transcription performed according to instructions in the Plant RNA maxi kit and EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMix, respectively
RT-qPCR analysis performed using TransStart Probe qPCR SuperMix and detected using a CFX96 Touch™ Real-Time PCR Detection System
Tubulin (accession No. AB239680.1) used as an internal reference gene

controls

Positive control: None mentioned
Negative control: None mentioned

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