http://www.openwetware.org/index.php?title=Cfrench:DNAextraction1&action=history Revision history for this page on the wiki en MediaWiki 1.13.2 Wed, 22 May 2013 19:41:34 GMT http://www.openwetware.org/index.php?title=Cfrench:DNAextraction1&diff=90711&oldid=prev <p></p> <p><b>New page</b></p><div>==Purifying DNA fragments using glass beads==<br /> '''From gel:'''<br /> <br /> *weigh gel slice<br /> <br /> *add 3 x weight in mg of 6 M NaI<br /> <br /> *melt in 55 C waterbath for about 5 minutes. Check and mix every 2 minutes or so. Make sure that gel has completely melted before next stage, but don't leave at 55 C any longer than necessary.<br /> <br /> '''From liquid:'''<br /> <br /> *add 3 x volume of 6 M NaI.<br /> <br /> '''In both cases:'''<br /> <br /> *add 5 ul glass bead suspension (make sure glass beads are well suspended before taking 5 ul). Mix and leave on ice for at least 10 minutes.<br /> <br /> *spin briefly (30 seconds to 1 minute). Remove supernatant and discard.<br /> <br /> *add 0.25 ml ice-cold wash buffer (stored in freezer). Respin, remove and discard supernatant.<br /> <br /> *repeat the previous step twice more (total of 3 washes). After the last wash, remove as much wash buffer as possible from the bead pellet.<br /> <br /> *resuspend pellet thoroughly (by pipetting up and down) in 15 ul of EB buffer.<br /> <br /> *return to 55 C waterbath for 10 minutes, mixing again (by flicking) after 5 minutes or so.<br /> <br /> *spin out glass beads and transfer supernatant (containing DNA) to a fresh tube.<br /> <br /> [[Cfrench:labtoppage|Back to main lab page]]</div> Thu, 07 Dec 2006 09:59:30 GMT Cfrench http://www.openwetware.org/wiki/Talk:Cfrench:DNAextraction1