DNA recovery from agarose gels-NA45

Domingos henrique D_henrique at icrf.icnet.uk
Tue Jul 19 05:05:33 EST 1994


About the NA-45 DNA recovery method I would suggest two small
modifications:
1. Add 1 microliter of glycogen (BMB or Appligene) to the eluate before
EtOH ppt and don't use any salt in this first EtOH ppt (is already 1M
NaCl). It helps in seeing the DNA pellet later and increases efficiency of
ppt. It has no effect in any subsequent treatment!!
2. If you want super-clean DNA do a phenol/CHCl3 extraction after the first
EtOH ppt and then reppt. Don't do phenol/CHCl3 extraction on the high salt
buffer as the DNA at this salt concentration (1 M salt) will remain mostly
at the interface. Don't need to add Glycogen again as it stays in the
aqueous phase.

And this is indeed the best method to purify DNA from gels.



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