A polymerase chain reaction-based DNA typing method, amplified fragment length polymorphism (AMP-FLP) analysis, has shown promise as a means of analyzing forensic biological evidence. A variety of DNA extraction methods were evaluated for their suitability for AMP-FLP analysis. Factors that were considered in the evaluation included DNA yield, ability of DNA to be amplified, the presence of DNA fragments other than those expected for the alleles in the sample, and differential amplification of different sized alleles for a sample. An initial screen of eight extraction methods was conducted on bloodstains deposited on cotton sheeting. These methods included Chelex | 100, organic extraction followed by Centricon 100 |
The interlaboratory typing of DNA specimens that have been amplified at the D1S80 locus necessitates the use of a standard allelic reference ladder. This communication describes a technique in which individual, amplified alleles are isolated, combined, and amplified by PCR to produce a functional reference ladder composed of many of the alleles that occur at this locus. The amplified ladder can serve directly as a template source for production of the next generation of reference ladder. This process, in which each amplified ladder serves as the template for the next has been carried through multiple generations.
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