I am using a commercially available kit for RNA Isolation to purify tRNA, however this kit is not suited for purification of short RNA. The supplier recommended that instead of using the supplied binding buffer I should add 0.1 volumes of NaOAc pH 5.2 and 3 volumes of isopropanol, the rest of the purification protocol is unchanged. This might work for you as well. If you don't want to "hijack" a commercially available kit, I recommend the following literature:

Boom, R., C. J. Sol, M. M. Salimans, C. L. Jansen, P. M. Wertheim-van

Dillen, and J. van der Noordaa. 1990. Rapid and simple method for purification

of nucleic acids. J. Clin. Microbiol. 28:495–503.

Boom, R., C. Sol, M. Beld, J. Weel, J. Goudsmit, and P. Wertheim-van

Dillen. 1999. Improved silica-guanidiniumthiocyanate DNA isolation procedure

based on selective binding of bovine alpha-casein to silica particles.

J. Clin. Microbiol. 37:615–619.

These publications describe a simple silica-based DNA-isolation method that actually works and you can prepare all buffers yourself.

Good luck!

Best regards,

John

well I had a problem with extraction DNA from biofilms and durind 3 months I tested as many protocols as it was possible.... Nevertheless one of the most important thing was to have pH of my samples almost 7.0 (from 6.6 to 7.0) and used NaOAc pH 7.3 and 0.7 volumes of isopropanol.

Maybe it would help in your reseach as well.

Best,

Tom

Hi all,

check the recipes of QIAGEN buffers at http://openwetware.org/wiki/Qiagen_Buffers

You can prepare your home made solution based on those.

cheers

Thanks to all!

I've choosed after all 4.2M GuHCl/isopropanol buffer for phages lysis and  silica spin adsorbtion and obtained ~250 ng/µl pure ssDNA

Hi Dmitri,

Would you tell me where your silica spin columns come from? Cause I've been selecting the high-quality column for 3 months, but I got no one suitable.