Summary:
Today, polymerase chain reaction is a common part of approaches serving for identification of
individuals in legal medicine. This method is easily practicable, however attention must be paid
to the optimization of reaction conditions and to the interpretation of results. From the literature,
such cases are known, in which during amplification of extremely small amount of DNA (e.g. from
one cell) the polymerase chain reaction preferably amplifies only one of two in the template DNA
present alleles. If the amplified fragments differ in length, the shorter one is amplified preferably,
and it may be cause of false results. In the presented study, DNA from 23 stains of male blood on
different fabrics was isolated by two different methods (by treatment with proteinase K and
boiling and by treatment with Chelex 100). The obtained DNA samples were amplified using
primers, they are complementary to the amelogenin gene sequences. The system is suitable for
sex determination, because amplification of the X-chromosomal sequence provides a fragment in
length of 632 bp, amplification of the Y-chromosomal one a fragment in length of 443 bp. The
isolation method based on proteinase K led in 17.38 % of samples to the very intensive preferential
amplification of the longer allele, and therefore to a false result. The isolation method based on
Chelex 100 provided in all cases correct results with clearly recognizable preferential amplificati-
on of the shorter allele. The reported results accentuate the meaning of choice of the appropriate
isolation method, the need of accurate PCR optimization, and the careful interpretations of its
outputs.
Key words:
polymerase chain reaction (PCR) - isolation of DNA - identification of individuals -
preferential amplification
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