Ultrasybr green mixture qpcr kit

Total RNA was isolated from Arabidopsis roots and shoots using a Total RNA Miniprep Kit (CWBIO, Beijing, China). cDNA synthesis was carried out using the RevertAid first-stand Synthesis System Kit (Thermo Fisher Scientific, Waltham, MA, United States). An UltraSYBR Green Mixture qPCR Kit (CWBIO) was used for the qPCR reaction, following the manufacturer’s protocol. Gene expression was determined by real-time PCR using an ABI7500 Fast Real-Time PCR System (Thermo Fisher Scientific). TUB2 (At5g62690) was used as the internal reference gene.

Total RNA was prepared from spines at different developmental stages using a RNAprep pure Plant Kit (TIANGEN, Beijing, China), according to the manufacturer’s instructions. The extracted RNA was treated with RNase-free DNase I (Fermentas, Harrington, Canada) to eliminate genomic DNA contamination, according to the manufacturer’s instructions. cDNA synthesis was carried out using a RevertAid First Strand Synthesis Kit (ThermoFisher Scientific). An UltraSYBR Green Mixture qPCR kit (CWBIO) was used in the qPCR reaction to determine the expression of relevant genes using an iCycler iQ™ multicolour real-time PCR detection system (Bio-Rad). Expression of genes was normalised to that of the cucumber actin gene. The primers used are listed in Supplementary Table S3 at JXB online.

Total RNA was extracted from plants using an ultrapure RNA kit (CWBIO). cDNA samples were carried out using a EasyScript One-Step gDNA Removal and cDNA Synthesis SuperMix Kit (TransGen Biotech). The UltraSYBR Green Mixture qPCR kit (CWBIO) was used in the qPCR reaction. The real-time PCR was carried out using VIIA7 (Applied Biosystems). We used TUB2 (At5g62690) as the internal control.