Hi all,
I've heard that SPRI beads also work for ssDNA, and I'd like to do a SPRI bead size selection of ssDNA > 150 nt (for various reasons I won't go into) but I'm a bit worried about the shorter ssDNAs hitching a ride on the longer fragments by non-specific base-pairing at room temperature. I'm using Ampure XP beads. So my question is, can these beads be used at elevated (stringent) temperatures, e.g. 55 - 65 degrees? Or could I use a denaturing agent like formamide to destabilize base-pairing?
After reading this thread: http://seqanswers.com/forums/showthread.php?t=31787 it seems to have been the consensus that even short ssDNA regions in the ladder can mess with the size selection. Any ideas / experiences to share?
I've heard that SPRI beads also work for ssDNA, and I'd like to do a SPRI bead size selection of ssDNA > 150 nt (for various reasons I won't go into) but I'm a bit worried about the shorter ssDNAs hitching a ride on the longer fragments by non-specific base-pairing at room temperature. I'm using Ampure XP beads. So my question is, can these beads be used at elevated (stringent) temperatures, e.g. 55 - 65 degrees? Or could I use a denaturing agent like formamide to destabilize base-pairing?
After reading this thread: http://seqanswers.com/forums/showthread.php?t=31787 it seems to have been the consensus that even short ssDNA regions in the ladder can mess with the size selection. Any ideas / experiences to share?
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