Protocol Online logo
Top : Forum Archives: : Molecular Biology

Help-How is the RNA quality? - (Feb/18/2008 )

From left to right:
1,2,3,4,DNA Marker,5,6,7,8
1,2: RNA extracted 40 days before, kept at -20 =C
3,4: 23 days before, kept at -20 =C
5,6: 13 days before, kept at -20 =C
7,8: 5 days before, kept at -20 =C

they were run on an agarose gel(1%), not denaturing agarose gel.
these RNA can be reverse transcribed?

Thank you!!!

-phhhhp-

Well, they look kind of degraded. Are you using RNAse-free buffers to run these samples? Are you sure all the stuff you're working with is RNAse free?

For long term storage keep your RNA at -80ºC. A couple of weeks at -20ºC won't be very problematic but -80ºC is always better.

-Ambrósio-

Thank you, Ambrósio.
Do you think the degradation is serious?
Because the Background is also high light, even the DNA Marker has smearing.
I don't use RNAse-free buffers to run the RNA, just common TBE buffer.
There are 3 phases these RNA may degrade, isolation of RNA, storage and running on agarose gel.
If I do the RT just after RNA isolation, do you think the cDNA is able to be subjected to Real-time PCR?

-phhhhp-

I just replied to your PM

-Ambrósio-