Detect gene in stable cell line, - (May/11/2009 )
I establish stable cell line expresses one scFv (VH-linker-VL). I check the expression of this gene in cell by RT-PCR. But i can only detect VH or VL (350 bp) without VH-linker-VL (750 bp). I change many kinds of primers. But i can't detect full scFv.
Would you please giving me some explanations? Thank you.
Size of full length product too big for the PCR?
No. It's only 750 bp.
oops, sorry should have read your post better
What about just PCR, do you have one piece VH-linker-VL? Could it be integrated and expressed as two pieces not a single VH-linker-VL? How many stable clones have you screened?