Trnzol a rna purification reagent

The roots of 3-week-old hydroponically cultured Col-0 and sic1 plants were used to extract total RNA by using TRNzol A+ RNA Purification reagent (DP421; Tiangen Biotech, Beijing, China). Two micrograms of total RNA were used to synthesize first-strand cDNA with TransScript one-step gDNA removal and cDNA synthesis super mix (AT311-02; TransGen Biotech, Beijing, China). qRT-PCR was performed using SYBR green PCR master mix (TRT-101; TOYOBO) with the first-strand cDNA as a template on a real-time PCR system (CFX thermocycler; Bio-Rad, ‎Hercules, CA). Primers for qRT-PCR were designed using Primer Express software version 3.0 (Applied Biosystems, Foster City, CA). The primers UBCF and UBCR were designed for ubiquitin-conjugating enzyme 21 (At5g25760), which was used as the reference gene. The primer sequences are shown in S8 Table. Expression data analysis was performed as previously described [51 (link)].

Total RNA was extracted from plants using TRNzol A+ RNA Purification reagent (TIANGEN, DP421, Beijing, China). Two micrograms of total RNA were used to synthesize first-strand cDNA with TransScript One-Step gDNA Removal and cDNA Synthesis SuperMix (Transgen, AT311-02, Beijing, China). qRT-PCR was performed using SYBR Green PCR Master Mix (TRT-101, TOYOBO, Osaka, Japan) with the first-strand cDNA as a template on a Real-Time PCR System (Bio-Rad CFX thermocycler, California, USA). Primers for qRT-PCR were designed using Primer Express Software Version 3.0 (Applied Biosystems, USA). The primers HKT1 exon2-3-1L and HKT1 exon2-3-1R were designed to span an exon-exon junction and were used to detect the gene expression level. The primers UBCF and UBCR were designed for UBIQUITIN-CONJUGATING ENZYME21 (At5g25760), which was used as the control gene. The primer sequences are shown in S3 Table. Expression data analysis was performed as described previously [34 (link)].