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 amplify small DNA fragments a billion-fold. The generation of amplicons either by PCR with a set of oligo primers or by arbitrarily primed AP-PCR with a single oligonucleotide primer is based on the availability of intact template and priming sites. With these approaches, it is possible to generate specific and random amplicons to assess the extent of damage to DNA caused by any of the physical, chemical, or environmental 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 that were boiled (treated) for different lengths of time. No difference was noticed in the amplification profile of DNA cooked for various lengths of time. However, after HinfI treatment, AP-PCR amplification of these DNAs revealed more bands on agarose gel than unrestricted samples. 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 of DNA probe technology is discussed.
Mohan
D
D
National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India.
Rao
K B
KB
Dixit
A
A
Ali
S
S
D000818:Animals; D001483:Base Sequence; D002020:Buffaloes / Q000235:genetics*; D004247:DNA / Q000032:analysis; D017931:DNA Primers / Q000235:genetics*; D005260:Female; D006358:Hot Temperature; D008297:Male; D008969:Molecular Sequence Data; D009132:Muscles / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D013048:Specimen Handling
D017931:DNA Primers; D004247:DNA