Total RNA isolation and cDNA synthesis were performed as explained in above section. We have used diluted cDNA (100 ng μL−1) for qRT-PCR reactions. Oligo’s for the qRT-PCR was designed using Genefisher2 software. We selected heat-responsive transcription factors - HSFA6e (acc. no. KU291394.1), WRKY (acc. no. KU562861.1), HSPs - HSP17 (acc. no. JN572711.1), HSP26 (acc. no. AF097659.1)], antioxidant enzymes – SOD (acc. no. AF092524.1), POX (acc. no. AF005087.1) and starch biosynthesis pathway linked genes – ADP-glucose pyrophosphorylase (large subunit; acc. no. KC347594.1), soluble starch synthase gene (acc. no. KJ854903.1) for the expression analysis (Table S3). The samples were used in triplicates for the expression analysis. The expression was performed following the protocol as mentioned in Kumar et al. [20]. We have used β-actin gene (acc. no. AB181991.1) for normalizing the Ct value. Further, Pfaffl method [24] was used for calculating the relative fold expression of the genes.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.