Rapid DNA Extraction from Dried Milk Spots: Application in Non-invasive Detection of the A1/A2 Variants of Beta Casein in Cow
2018
Venkatesh, Premakumari | Gopal, Dhinakarraj
Bovine’s milk protein constitutes around 30 % of beta casein (β-casein). Numerous studies have focused on the detection of the A1 variant of β-casein, as it often manifests with gastrointestinal complications. In the present study, dried milk spot (DMS) was used for sample collection, DNA extraction and for the detection of A1/A2 variants of β-casein in milk sample using polymerase chain reaction (PCR). Milk samples and blood samples were collected in sterile tubes from healthy cows at University Research Farm, TANUVAS, Chennai. DNA was efficiently extracted from DMS and dried blood spots (DBS) in nuclease free water by incubating at 95 °C for 5 min and PCR was performed for diagnostic screening of A1/A2 variants of β-casein. PCR analysis of milk samples collected on DMS correlated 100 % with those of DBS collected on FTA cards. PCR for A1/A2 genotyping yielded the expected amplicon size of 244 bp for all the samples. From the present experimental study, it is evident that DMS could be stored for 1 month at room temperature for diagnostic purposes. The current technique will emerge as a suitable non-invasive diagnostic method with minimal sample utilization and can be applied for screening various genetic disorders.
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