Detection of very-rare-copy DNA in 0.2-ml dried human blood blots.

摘要: A nested PCR assay for chromosome 7 inversions (as identified by the presence of T-cell receptor trans-rearrangements) can detect as rare a frequency 1 copy in 300,000 leukocytes. To identify such occurrences from dried blood blots, most conveniently obtained and stored samples field population studies, demands DNA extraction method that will provide both high quality yield. We have satisfied this requirement extracting proteins other components directly minced filter with phenol, before Chelex-water. This provides near maximal yield denatured sufficient to these translocations sensitivity equivalent purified whole samples. Blots 6 months worked well fresh blots. In addition, we present obtaining native although at much lower The successful use blots events signals feasibility large-scale studies involving diagnostic molecular epidemiology.