Small RNA - FlashPAGE

[RickC7]

Hi Everyone,

Has anyone tried using the FlashPAGE system to do library size selection from a finished small RNA library? Using the SOLiD protocol this would mean recovering the 105-150bp size range. Just wondering about the time settings and any helpful hints.

Many Thanks,
Richard

[scooter]

It is typically hard to calibrate flashpage to give you the tight cut-off needed for the particular size range you need. I am assuming you want to purify away the adaptor only products from the products containing the RNA sequences of interest?

This can be done, however the instrument wasn't really designed to run for extended periods of time and salt depletion can cause your power supply to conk out if it cannot handle very low mA readings. Otherwise, flashpage is a wonderful tool.

[jinzhuuiuc]

Isn't Flashpage designed for separating single stranded nucleic acid? Would the ds DNA library run safe and sound through the gel?

[scooter]

FlashPAGE gels are denaturing conditions and is indeed designed for single-stranded RNA, so your DNA would probably not stay double-stranded.