No cDNA from RNA sample - (Aug/25/2008 )
Hi,
I have isolated RNA from barley tissue using a phenol/chloroform/Lithium Chloride method. The RNA was then DNAsed using the turbo dnase kit
the RNA looks good on a formaledehyde gel and also by nanodrop and spectrophotometry the amounts are good (50-60 ng/ul)
I sent this RNA for microarray analysis but the samples couldn't be detected in their bioanalyzer. They put the RNA though the spec and it appeared fine. I have since tried converting to cDNA but did not get and DNA
Could DEPC be a possible inhibitor?
Thanks
No, DEPC will not affect the result. YOu saw sharp bands on the gel when you checked?
yes bands were quite sharp, with little or no degradation
Autoclave DEPC treated water before usage, I think you have done.
YOu could try to change the tubes and water you have used. One of my friends had the same experience as you, when he changed those, all problems were fixed.
yes bands were quite sharp, with little or no degradation
[quote name='microlight' date='Aug 25 2008, 03:57 PM' post='148330']
Autoclave DEPC treated water before usage, I think you have done.
YOu could try to change the tubes and water you have used. One of my friends had the same experience as you, when he changed those, all problems were fixed.
have autoclaved water.
will try to change water and tubes
thanks