Degraded total RNA - (May/08/2005 )
What happens if my total RNA is degraded? Should i use it further in my work? 
hi
it depends if you can see the bands of rna "inside the smear" of degraded rna. I've use partially degraded rna and orks fine. I used it on acrylamide gels and usually you can see many bands on it. There was a smear but not that intense and i was able to saw the bands in it.
Can you send a photo of your gel?
it depends if you can see the bands of rna "inside the smear" of degraded rna. I've use partially degraded rna and orks fine. I used it on acrylamide gels and usually you can see many bands on it. There was a smear but not that intense and i was able to saw the bands in it.
Can you send a photo of your gel?
Thanks, fred. I can't take the picture, i don't have the equipment to do it.
I can't see the bands, but would this hurt if i use this sample for real time PCR or should i disgard it and start a new isolation?
hum
well if you see the bands that mean your rna is quite good. try your rtpcr, it's a very sensitive technique so i assume you would get your answer if you only check presence. If it's for quantitative rtpcr, i've never done it, but i would say an other solation may be better
Hi!
To see the quality of my RNA I always use a formaldehid gel. To know is your RNA are good for real time PCR or to make RTPCR, you have to see the two ribosomic bands, 18 and 16S. When your RNA is degradated you can see a big smear or the 18S less intense than you 16S, it means is degradated, you have to use RNA that have a intense bands.
I'll hope this can help you to choose your RNA.
Good luck!
baobab
Khmmm... If possible, prepare a new RNA sample. Otherwise, it could be GIGO.
If you have a 28S RNA band, I would say, give it a shot. Otherwise, just forget about it! The result will not be reliable. BTW, did you figure out why your RNA is degraded? Is it from tumor tissue or a cell ine?
SV