microarray confirmation - which method? (Jan/23/2007 )
hi, I have just done some microarrays and Im just wondering what is the most accepted way of confirmation of data? Is semi-quantitative PCR acceptable? or does it have to be real time PCR? Or could you use protein? semi quant would be the most convenient as we have everything needed in the lab. Plus I really suck at real time PCR any thoughts welcome
I think the best method of verification is regular RT-PCR. Only when RT-PCR cannot tell a difference, do people turn to real-time which can pick up small differences.
i always do a RTPCR before attempting real time. a, there is something see on a gel, instead of looking at a computer screen, b, you know that the primers et al are working.
In case you work with cell cultures it would be RT PCR. In case you isolated tissue/cells from tissue then it would be ISH.