Extraction from Chitin - (Dec/08/2005 )
I am trying to extract DNA from insect wings using Chelex (had no luck with penol/chloro) and find my DNA/RNA ratios to hover in the 1.0 to 1.4 range. 
Any ideas on how to either reduce the RNA or increase the DNA ratio?
-bigbluebird-
its better to check the quality of DNA on agarose gel as well...
compare the amount with DNA marker, if its ok u can take it furthr for PCR.
other wise only properly done phenol:chloroform extraction is the best solution to my knowledge till now
check ur phenol also as at low pH ie around 4.0 instead of 8 its not possible to have good quality of DNA
good luck
-cherub-
QUOTE (bigbluebird @ Dec 8 2005, 09:39 PM)
I am trying to extract DNA from insect wings using Chelex (had no luck with penol/chloro) and find my DNA/RNA ratios to hover in the 1.0 to 1.4 range. Any ideas on how to either reduce the RNA or increase the DNA ratio?
Any ideas on how to either reduce the RNA or increase the DNA ratio?-bigbluebird-
QUOTE (cherub @ Dec 9 2005, 04:21 AM)
its better to check the quality of DNA on agarose gel as well...
compare the amount with DNA marker, if its ok u can take it furthr for PCR.
other wise only properly done phenol:chloroform extraction is the best solution to my knowledge till now
check ur phenol also as at low pH ie around 4.0 instead of 8 its not possible to have good quality of DNA
good luck
compare the amount with DNA marker, if its ok u can take it furthr for PCR.
other wise only properly done phenol:chloroform extraction is the best solution to my knowledge till now
check ur phenol also as at low pH ie around 4.0 instead of 8 its not possible to have good quality of DNA
good luck
Thanks. I'll run a gel on Monday. Off to snowland in the morning. (I think that's allowed since there are no dragonflies flying in the northeast right now.)
The protocol for insect wings is Chelex, but I'll try chloro/phenol on Monday, too.
-bigbluebird-