OCT medium and real time pcr - (Nov/04/2005 )
Has anyone got any experiences with real time PCR using RNA converted to cDNA that is extracted from tissues embeded in OCT medium?
We are trying to do that in our laboratory but real-time PCR seems to be inhibited!! Previously, using the same Taqman assays, we were able to see amplifications from cDNA obtained from freshly cultured cell lines. However, now that we switched to tissues embedded in OCT, we cannot get any amplification.
Does any one have similar experiences? How can we go around this problem?
How did you treat you sample before you embedded it into OCT medium?
Fixed in aldehydes before freezing them?
How did you extract your RNA?
The samples are freshly embedded into OCT medium. Not fixed at all in aldehyde.
RNA extraction was done using Qiagen kits.