Cells, grown in selective SD media (20 ​ml in 100 ​ml flasks) at 30 ​°C and 200 ​rpm until mid-log phase, were harvested and total RNA was isolated using HiPurA Yeast RNA Purification Kit (HiMedia, India-MB611). The 100 ​ng of isolated RNA was used to prepare cDNA using a cDNA synthesis kit (BIO-RED Cat#170–8891). For quantitative Real-Time PCR (qRT-PCR), cDNA (1 ​μl) was amplified using iTaq™ Universal SYBR® Green Supermix qPCR kit (BIO-RED, CAT#172–5124) by using eGFP specific primer (Table S1) and following manufacturer’s protocol. The reaction was subjected to an initial denaturation at 95 ​°C for 2 ​min and 40 repetitive cycles of 15 ​s ​at 95 ​°C, 30 ​s ​at 60 ​°C in CFX96 Touch Real-Time PCR System (BIO-RED). The 18 ​S rRNA was selected as a reference gene.

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.