Rotor gene 6000 instrument system

Total RNA was isolated from the samples of various tissue types using RNAiso Plus (TaKaRa, Japan; http://www.takarabio.com), and was quantified by a Nano Nanodrop ND-2000 Spectrophotometer (Thermo Scientific, USA, http://www.nanodrop.com), as described previously (Cho et al., 2018 (link)). The cDNA was prepared with 10 ng of the oligo (dT)₁₈ primer, 2.5 mM deoxy ribonucleotide triphosphates, and Moloney murine leukemia virus reverse transcriptase. Quantitative real-time RT-PCR (qRT-PCR) was performed with the synthesized cDNAs as templates and the gene-specific primers (Supplementary Table S1), using SYBR Premix Ex Taq™ II (TaKaRa) and the Rotor-Gene 6000 instrument system (Corbett Research, Sydney, Australia; http://www.corbettlifescience.com). Rice ubiquitin 1 (Ubi1) served as an internal control, and at least three biological replicates were analyzed. We used the data only when the melting curve showed a single sharp peak.

Leaf blades and root tissues were harvested at 7, 14, 21, 28, and 35 DAG from SD-grown ‘Longjing27’ plants. Total RNA was isolated from the samples using RNAiso Plus (TaKaRa, Shiga, Japan; http://www.takarabio.com) and qualified by a Nano Nanodrop ND-2000 Spectrophotometer (Thermo Scientific, Wilmington, DE, USA; http://www.nanodrop.com) as described previously (Cho et al., 2016 (link)). Complementary DNA (cDNA) was made with 10 ng of the oligo (dT)₁₈ primer, 2.5 mM deoxy ribonucleotide triphosphates, and Moloney murine leukemia virus reverse transcriptase. Quantitative real-time RT-PCR (qRT-PCR) was performed with the synthesized cDNAs as templates and primers listed in Supplementary Table S1, using SYBR Premix Ex Taq™ II (TaKaRa) and the Rotor-Gene 6000 instrument system (Corbett Research, Sydney, Australia; http://www.corbettlifescience.com). Rice Ubi1 served as an internal control. At least three biological replicates were analyzed. We used the data only when the melting curve showed a single sharp peak.