Questions about single pig oocyte RT-PCR.

[dragonwang]

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!
My email: [email protected]

[varshaparekh]

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