Introduction to basic mouse handling techniques.
Purification of DNA from fixed, paraffin-embedded tissue (PET) requires the removal of paraffin, proteins, and, in some cases, heavy metal ions. The resulting DNA is not generally suitable for Southern blot analysis. However, the polymerase chain reaction (PCR) can be used to amplify short regions of this DNA. In this appendix, the basic protocol can be performed rapidly and involves only a small number of manipulations, thus minimizing the possibility of contamination. An alternate protocol involves additional steps including extractions with xylenes and phenol/chloroform. This procedure is recommended as the primary approach when tissue has not been fixed in an optimal manner or as a secondary approach when the basic protocol has failed to produce satisfactory DNA.
Donovan
John
J
Rhone-Poulenc Rorer, Collegeville, Pennsylvania, USA.
Brown
Patricia
P
Reeves
J P
JP
Reeves
P A
PA
D000818:Animals; D000830:Animals, Laboratory; D015139:Blotting, Southern; D006224:Cricetinae; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D051379:Mice; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D051381:Rats
D004247:DNA