Optimisation of DNA and RNA extraction from archival formalin-fixed tissue.
Archival, formalin-fixed, paraffin-embedded tissue is an invaluable resource for molecular genetic studies but the extraction of high quality nucleic acid may be problematic. We have optimised DNA extraction by comparing 10 protocols, including a commercially available kit and a novel method that utilises a thermal cycler. The thermal cycler and Chelex-100 extraction method yielded DNA capable of amplification by PCR from every block and 61% of sections versus 54% using microwave and Chelex-100, 15% with classical xylene-based extraction and 60% of sections using the kit. Successful RNA extraction was observed, by beta-actin amplification, in 83.7% sections for samples treated by the thermal cycler and Chelex-100 method. Thermal cycler and Chelex-100 extraction of nucleic acid is reliable, quick and inexpensive.
Coombs
N J
NJ
University Surgical Unit, Southampton General Hospital, Southampton SO9 4YD, UK.
Gough
A C
AC
Primrose
J N
JN
D001109:Archives; D004247:DNA / Q000302:isolation & purification*; D005557:Formaldehyde; D016612:Paraffin Embedding; D011309:Preservation, Biological; D012313:RNA / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D016707:Tissue Fixation
D005557:Formaldehyde; D012313:RNA; D004247:DNA