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Questions about single pig oocyte RT-PCR.

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Questions about single pig oocyte RT-PCR.

Postby dragonwang » Mon Jun 23, 2008 3:48 am

My lysis buffer:
NP40 2ul
RNase Inhibitor 5ul
dNTP 10ul
oligodT 10ul
DTT 10ul
5*RT buffer 40ul
Add H2O to 100ul,
Take 5ul to eppendorf tube.

hyaluronidase to remove cumulus cells.
The amplification program for the reverse transcription step was as follows:
70°C for 10 min, ice 2min ,MMLV ,42°C for 50min, 95°C for 5 min.
then PCR.
Why no result???
I am puzzling! :cry:
My email: [email protected]
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Joined: Mon Jun 23, 2008 3:42 am

Postby varshaparekh » Sat Jul 12, 2008 3:58 pm

may be ur lysis buffer is not compatible. add some DTT to it
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