Easypure plant rna kit

Total RNA from poplar samples was isolated using PureLink Plant RNA Reagent (Invitrogen), and cleaned with RNeasy Plant Mini Kit (Qiagen) as described previously (Geraldes et al., 2011 (link); Wang et al., 2014 (link)). Total RNA from Arabidopsis seedlings was isolated using EasyPure^TM Plant RNA Kit (Transgene) according to the manufacturer’s instructions. All RNA samples were treated with RNase-Free DNase set (Qiagen) to eliminate possible DNA contamination.
cDNA was synthesized using 2 μg total RNA by Oligo(dT)-primed reverse transcription using Omniscript RT Kit (Qiagen). Some of the primers used for cloning or examining the expression of corresponding genes have been described previously (Wang et al., 2007 (link), 2008 (link), 2010 (link); Gan et al., 2011 (link)), and poplar R3 MYB gene-specific primers are shown in Table 1.

Ten-day-old Arabidopsis and rice seedlings were used for RNA isolation. Total RNA was isolated by using an EasyPure^TM Plant RNA Kit (Transgene Biotech) and following the manufacturer’s instructions. cDNA was synthesized by using an EasyScript First-Strand DNA Synthesis Super Mix (TransGen Biotech) and following the manufacturer’s procedures, and used for RT-PCR and qRT-PCR amplification. For expression analysis of TTG1 and OsTTG1A, the expression of ACTIN2 (ACT2) was used as a control. For expression of GL2 and R3 MYB genes, the expression of ACT2 was used as an inner control.
The primers used for amplification of OsGL1A are 5’-CAACATATGATGGGGAGGTCGCCGTGC-3’and 5’-CAACTTAAGTCATTTCATGGGGAGGCTTCTG-3’, for OsGL1B are 5’-CAACATATGATGGGGAGGTCACCG-3’and 5’-CAACTTAAGTCATTTCATTTCCAAGCTTCTG-3’, for OsTTG1A are 5’-CAACATATGGAGCAGCCCAAGCCG-3’ and 5’-CAACTTAAGTCAGACCCTGAGAAGCTGGA-3’, for GL1 are 5’-CAACATATGAGAATAAGGAGAAGAGATGA-3’ and 5’-CAACTTAAGCTAAAGGCAGTACTCAACATC-3’, for TTG1 are 5’-CAACATATGATGGATAATTCAGCTCCAGATTCG-3’ and 5’-CAACTTAAGTCAAACTCTAAGGAGCTGCATTTTG-3’, and for GL3 are 5’-CAACATATGGCTACCGGACAAAACAG-3’ and 5’-CAAGAGCTCTCAACAGATCCATGCAACCC. Other primers used for RT-PCR have been described previously [40 (link), 48 (link)].

Total RNA from rice was isolated as described previously for RNA isolation from poplar48 (link)49 50 (link). Total RNA from Arabidopsis seedlings was isolated using EasyPure^TM Plant RNA Kit (Transgene Biotech) according to the manufacturer’s instructions.
cDNA was synthesized using total RNA isolated by Oligo(dT)-primed reverse transcription using EazyScript First-Strand DNA Synthesis Super Mix (TransGen Biotech) following the manufacturer’s procedures. qRT-PCR was performed on the Applied Biosystems 7500 real time PCR System using SYBR Green/ROX Master Mix (Thermo Scientific). The primers used for qRT-PCR examination of TCL1, TRY, CPC, GL1, GL2, TUBULIN2 and OsUBQ5 have been described previously26 (link)51 (link)52 (link). The primers for other Arabidopsis and rice genes are listed in Table 1.