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Detect gene in stable cell line,


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#1 Pham Dinh Chuong

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Posted 11 May 2009 - 06:15 PM

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.

#2 bob1

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Posted 12 May 2009 - 05:43 PM

Size of full length product too big for the PCR?

#3 Pham Dinh Chuong

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Posted 14 May 2009 - 02:38 AM

No. It's only 750 bp.

#4 bob1

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Posted 14 May 2009 - 04:15 PM

oops, sorry should have read your post better

#5 AquaPlasmid

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Posted 14 May 2009 - 10:10 PM

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?




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