Detect gene in stable cell line, - (May/11/2009 )
Hi,
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.
-Pham Dinh Chuong-
Size of full length product too big for the PCR?
-bob1-
No. It's only 750 bp.
-Pham Dinh Chuong-
oops, sorry should have read your post better
-bob1-
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?
-AquaPlasmid-