Thermal cycler dice real time system 2 tp800

Total RNA was extracted from each tissue of O/E and WT plants using an RNeasy Plant Mini Kit (Qiagen, Limburg, The Netherlands). Reverse transcription reactions and subsequent real time PCR analyses were performed using PrimeScript™ RT Master Mix and a Thermal Cycler Dice Real Time System II TP800 (Takara, Japan), as described previously [23 (link)]. Primer sequences for OsHKT1;4 and an internal control OsSMT3 were shown in Table S1.

Total RNA was extracted using an RNeasy Plant Mini Kit (Qiagen, Limburg, Netherlands), and reverse transcription was performed using PrimeScript™ RT Master Mix in accordance with the manufacturer’s protocol (Takara, Japan). A Thermal Cycler Dice Real Time System II TP800 (Takara) was used for the real-time PCR analysis. The Q-PCR analysis was performed setting the maximum cycle number of 40 in accordance with the protocol of the manufacturer (Takara, Japan). OsSMT3, HistoneH3 or Actin genes were used as internal controls as described previously [50 (link)]. Gene-specific primers for OsHKT1;4 were used: Forward, 5’-GTCGAAGTTGTCAGTGCATATGG-3’; Reverse, 5’-TGAGCCTCCCAAAGAACATCAC-3’.
To analyze the tissue specific expression of OsHKT1;4 in node I, regions of the enlarged vascular bundle (EVB) and the diffuse vascular bundle (DVB) were excised by the laser microdissection (LMD: Arcturus Laser Capture Microdissection System, Life Technologies, CA, USA) as described previously [53 (link)].