ASSESSMENT OF AMPLICONS IN THE DNA FROM BOILED TISSUE BY PCR AND AP-PCR AMPLIFICATION

Polymerase chain reaction (PCR) is a technique sensitive enough to amp lify small DNA fragments a billion-fold. The generation of amplicons e ither by PCR with a set of oligo primers or by arbitrarily primed AP-P CR with a single oligonucleotide primer is based on the availability o f intact template and priming sites. With these approaches, it is poss ible to generate specific and random amplicons to assess the extent of damage to DNA caused by any of the physical, chemical, or environment al factors. We report the amplification of sex chromosome and autosome specific loci in the buffalo (Bubalus bubalis) genome by symmetrical and AP-PCR performed on DNA samples isolated from the muscle tissues t hat were boiled (treated) for different lengths of rime. No difference was noticed in the amplification profile of DNA cooked for various le ngths of time. However, after HinfI treatment, AP-PCR amplification of these DNAs revealed more bands on agarose gel than unrestricted sampl es. The successful amplification of the DNA samples isolated from the boiled tissues is attributed to the intactness of the amplicons. This suggests that despite storage for more than a year and subsequent heat treatment to the muscle tissues, the DNA remains a good substrate for PCR and AP-PCR amplification. Relevance of this work in the context o f DNA probe technology is discussed.