the centrifuge speed to precipitate the 30bp oligo - (Jan/31/2006 )
i need to precipitate a 30bp length oligonucleotides by using glycogen as a carrier and ethonal. i dont know wether a centrifuge speed of 15000rpm is enough or not. can anyone tell me. thank you very much in advance.
i need to precipitate a 30bp length oligonucleotides by using glycogen as a carrier and ethonal. i dont know wether a centrifuge speed of 15000rpm is enough or not. can anyone tell me. thank you very much in advance.
Dear friend,
Using glycogen is a good idea. Perhaps you can try linearised acrylamide as well.
If you can't precipitate you 30bp oligo at maximum speed try spin longer period or use buthanol instate of ethanol.
Good luck. Please let us know if this work.
Hadrian
0.3M NaCl and 2 times ethonal plus glycogen, after >1 hour -70C
centrifuge at 4C for 30mins, spin longer time is better
Um, G for spin speed? Isn't that the most important thing here?
-Matt
in the paper "Profiling miRNA expression..." from Sioud et al., they allow 24h precipitation for small RNAs : briefly, total RNAs are isolated (using the guanidinium thiocyanate phenol-chloroform method (14), but I’ve done it with Trizol...) To precipitate the high molecular weight RNA, a 10% final concentration of polyethylene glycol (8000) and 0.5 M NaCl were added. The mixture was incubated at 4°C for 30 min. After centrifugation at1000× g at 4°C, low molecular weight RNA was precipitated from the supernatant by the addition of 3 volumes of ethanol for 24 h at -70°C. RNA pellets were washed with 70% ethanol, air dried, and then dissolved in DEPC-treated water. Small RNAs (16–26 nucleotides in length) were (PAGE)-purified and then dissolved in 20 μL DEPC-treated water. From 200 μg total RNA, [they] obtained enough material to prepare 3 cDNA probes.
a paper may interest you at this adress :
http://nar.oxfordjournals.org/cgi/reprint/34/2/e9
it's a free access article
i need to precipitate a 30bp length oligonucleotides by using glycogen as a carrier and ethonal. i dont know wether a centrifuge speed of 15000rpm is enough or not. can anyone tell me. thank you very much in advance.
Dear friend,
Using glycogen is a good idea. Perhaps you can try linearised acrylamide as well.
If you can't precipitate you 30bp oligo at maximum speed try spin longer period or use buthanol instate of ethanol.
Good luck. Please let us know if this work.
Hadrian
with glycogen , i centrifuged the DNA at 15000rpm, i did not lose my DNA, so this method works well. but my ligation could not work, do you have any suggestion? u can go to read my another post.