identifying isoforms - (Jun/17/2009 )
I'm new to thsi area of researh and would appreciate all advice. I need to look for isforms of a partiur gene in my panel of cancer tissues (T and NT). What is teh ebst way to approachthis? I was planning to design priemrs for full length CDS of my gene of interest and check the expression, and clone the band to confirm the sequence?
Is the isoform known and you just want to check its expression in your samples? If yes, you just need to design primers that differentiate the isoform from other isoforms. If you don't have any idea about the structure of the isoform, there are many strategies to study it. For example, Northern blotting by designing a probe on a region common to all isoforms. you can also clone the full-length cDNA for sequencing.