cDNA storing
 

Hi everybody

I have problems with cDNA storing, i have tried -70 and -20, but when i´m gonna work with this samples i don´t get any results...

I need help..!!!

Re: cDNA storing
 

Hi there
what are you trying to do with your cDNA?

You may have to look at how you are storing it:

A) temperature
B) Buffer and salts used as final storing - will this affect your enzymatic reactions?
ie your cDNA could be there but you cannot get anything from it
C) is your cDNA degraded or poor quality to begin with?

Re: cDNA storing
 

You should try working with your cDNA right away to make sure it IS there. Once you're certain that you succesfully obtained cDNA, you should test the storage conditions. Since you should have the resulting cDNA diluted on DEPC treated water (or something nuclease free), and since it is DNA, it should be safe stored at -20ºC, don't know how much time though.

Re: cDNA storing
 

Maybe its not bout the storage, perhaps its reverse transcription problem.
you should do a quality check on your cDNA right after RT, simply by conducting a RT-PCR with housekeeping genes.