Total RNA was isolated from cells with the RNeasy Mini Kit (Qiagen, 74106). Complementary DNA (cDNA) was synthesized with PrimeScript Reverse Transcriptase (TakaraBio, RR047A) according to the manufacturer’s protocol. All quantitative real-time PCRs (qRT-PCRs) were performed with a ViiA 7 Real-Time PCR System and Quanti Studio 3 (Applied Biosystems) using PowerUp SYBR Green Master Mix (Applied Biosystems, A25742) and custom-designed primers in a final volume of 10 μl. Cycling conditions were 98°C for 2 min, followed by 40 cycles of 98°C for 10 s and 60°C for 30 s. Relative expression of target genes was measured using the comparative ΔΔCt method, and human ACTB was used as internal control. The sequences of the qRT-PCR primers are listed in table S1.

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

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.