DNA extraction protocol for use on Crustaceans..?

Ed Rybicki ED at micro.uct.ac.za
Fri Jul 2 02:37:24 EST 1993


> Freeze, thaw, lyse in SDS buffer, add KOAC, cool, then phenol,
> chloroform-isoamyl, chloroform extractions, isopropanol pptn, cold EtOH wash,
> wick dry, parafilm overnight, resuspend in TE.

We managed to clean things up remarkably well (for PCR, or other NA
manipulations) by simply putting total plant nucleic acid preps (from
plants dripping with slime) thru a Sephadex G-100 mini-column (recycled 10
ml glass pipette): gets rid of lots of unspecified nasties which do evil
things to restriction enzymes, reverse transcriptase, Taq pol, etc.

  ____________________________________________________________________
 | Ed Rybicki, PhD             |       "Lord, won't you buy me        |
 | (ed at micro.uct.ac.za)        |                                      |
 | Dept Microbiology           |         A Mer-ce-des Benz..."        |
 | University of Cape Town     |                                      |
 | Private Bag, Rondebosch     |                                      |
 | 7700, South Africa          |           - Janis Joplin             |
 | fax: 27-21-650 4023         |                                      |
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