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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-