High-throughput DNA extraction from old paraffin-embedded biopsies.
We have developed a rapid and reliable microporous spin-column-based method for high-throughput extraction of DNA from old paraffin-embedded biopsies. The method was developed specifically for PCRs and molecular genetic studies. De-waxing and DNA extraction from tissues is performed in only one vessel, minimizing pipetting steps and eliminating supernatant/pellet procedures. All steps are performed at 60 degrees, 55 degrees and 95 degrees C minimize deleterious DNase activity. The amount of the extracted DNA was measured spectrophotometrically, and the quality of the extracted DNA and the PCR products was controlled on ethidium bromide-stained 3% agarose gel. The presented method for DNA extraction is less laborious and more convenient than previously published methods. It provides means by which extensive, paraffin-embedded material can be subjected to molecular genetic studies.
Jalouli
J
J
Sahlgrenska University Hospital, Ostra, Göteborg.
Sand
L
L
Gustavsson
B
B
Hirsch
J M
JM
Larsson
P A
PA
D000199:Actins / Q000235:genetics; D001706:Biopsy; D001709:Biotechnology / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D017931:DNA Primers; D006801:Humans; D010232:Paraffin; D016133:Polymerase Chain Reaction
D000199:Actins; D017931:DNA Primers; D010232:Paraffin; D004247:DNA