positive control for RT PCR - (Nov/09/2005 )
I am wanting to include a figure for a paper demonstrating the expression of receptors in my cell model, or the lack there of, in some cases. I was wanting to get some opinions as to whether it is ok to use genomic DNA as a positive control for the PCR conditions given that they are single exon genes? I will also include a RTPCR for a house keeping gene to demonstrate the quality of cDNA template? The problem is that usually the gene is only expressed in brain, and getting human brain tissue or cell lines is an issue given deadlines.
I think so, what I see as a possible problem is that single exon RT-PCRs will amplify gDNA contaminating your RNA prep... be sure to include no RT controls to show your prep is clean.
Thanks for that, i have DNase treated the preps and also included a negative, it shows no or very little expression, hence the need for a positive control. Thanks for your advice!!!