Sex identification of meat by Amelogenin XY gene
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Identification of sex of meat animals is an important issue as cow slaughter is
banned in India but not bullock slaughter. Keeping this in mind a method for identification
of sex of the animal from which the meat is produced has been standardized which
involves PCR amplification of Amelogenin XY gene. In males amino acid coding region
of the amelogenin gene on the Y chromosome (AMELY) is shorter than its X chromosome
counterpart i.e. it is having a deletion region. Hence PCR amplification of AMEL
gene gives amplicons of two sizes (217 & 280 bp) in male and only amplicon of single
size (280 bp) in female DNA samples (Fig 1). Repeatability of the assay was confirmed
by testing in different field samples.
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| Fig 1. PCR amplification products of DNA from cattle and buffalo meat |
| Line 1: Negative control | Line 2: Cattle (male) |
| Line 3: Cattle (female ) | Line 4: Buffalo (male) |
| Line 5: Buffalo (female) | Line 6: Marker |
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Identification of sex of meat by Multiplex PCR
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Sex of meat could be identified by PCR amplification of SRY gene. Positive PCR amplification
indicates that sample is of male and negative PCR amplification indicates the sample
is of female. The method is applicable for cattle and buffalo meat. In order to
overcome the problem of false negative results due to PCR failure, mitochondrial
12S rRNA primers were also used in the PCR reaction mix as positive control. PCR
amplification yielded amplicons of size 456 bp and 195 bp in male samples. Whereas,
in female sample, PCR yielded amplicon of size 456 bp only (Fig 1). In order to
complete the analysis in shortest time DNA extracted from Alkaline lysis method
was also used to multiplex PCR amplification of SRY gene and mt 12S rRNA gene. Results
were reliable and could be of use to identify sex of meat. It is having major application
in detection of cow meat.
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| Figure 1: Multiplex PCR for identification of sex of meat |
| CM: Cattle Male (Two amplicons – 456 bp and 195 bp) |
| CF: Cattle Female (Single amplicon of size 456 bp) |
| BM: Buffalo Male (Two amplicons – 456 bp and 195 bp) |
| BF: Buffalo Female (Single amplicon of size 456 bp) |
| C: Negative control; L: 100 bp Ladder |
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