Copy number analysis was performed using 2 μL of dilute DNA (5 ng/μL), 5 μL of Taqman universal PCR master mix, 0.5 μL of Taqman Copy Number Assay probe (FAM), 0.5 μL of Taqman Copy Number Reference Assay (RNaseP) probe (VIC) and 2.0 μL of RNase‐Free Water to produce a 10 μL reaction volume for analysis by an ABI ViiA7 real‐time PCR system (Thermo Fisher Scientific Inc). Gene copy number was normalized to the Ribonuclease P RNA component H1 (H1RNA) gene (RPPH1) on chromosome 14 and calculated using the ViiA7 software as comparative CT (ΔΔCT) values. The nonimmortalized normal kidney cell line, RPTEC, was designated to represent a normal chromosomal copy number with a value of 2. Taqman Copy Number Assay probes (Thermo Fisher Scientific Inc) were used to assess the gene copy number of the following genes: CDKN2A (Hs03714372_cn), MET (Hs01432482_cn), and EGFR (Hs00711969_cn).

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.