Rna prep pure kit

Total rice RNA was extracted with an RNA Prep Pure Kit (Zymo Research) according to the manufacturer’s instructions. First-strand cDNA was synthesized from 2 μg of total RNA using a SuperScript II Kit (TaKaRa). Primer pairs used for qPCR are listed in Supplementary Table S1 at JXB online. qPCR analysis was conducted using an ABI7500 fast qPCR system with the SYBR Premix Ex Taq (TaKaRa; RR041A). The procedure was as follows: initial polymerase activation for 30 s at 95 °C followed by 40 cycles of 95 °C for 5 s and 60 °C for 34 s. For each sample, qPCR was performed with three technical replicates on three biological replicates. The 2^–ΔΔCT method was used to analyse relative transcript levels in gene expression (Livak and Schmittgen, 2001 (link)). Primers used for ChIP-PCR are listed in Supplementary Table S1. For detecting the expression level of LP2 in the dst mutant, 4-week-old plants were used for RNA isolation.

Total RNA was extracted using an RNA Prep Pure Kit (Zymo Research, Orange, CA, USA). A reverse transcription (RT) kit (Qiagen) was used in RT reactions. Quantitative RT-PCR (qRT-PCR) was performed using SYBR Premix Ex Taq Kit (TaKaRa; RR041A) in an ABI PRISM 7900HT (Applied Biosystems), and ubiquitin was used as the internal control. Standard errors were calculated from three biological replicates.