Method of determination allelic variants of kappa casein and beta-lactoglobulin in cattle
Year | 2012 |
Cathegory | Patents |
Internal link | 12038.pdf |
Abstract | The solution relates to a determination of allelic variants of kappa casein and beta-lactoglobulin in cattle with touch down multiplex polymerase chain reaction (PCR). It is characterized by amplification of allelic variants performed with primers on the following composition: Kappa casein CSN3 F: 5 ‚- CCA TTG CTA GTG GTG CTA AGC C – 3 ‚(22 mer) and CSN3 R: 5‘ – CCT GCG TCT TCT TTG TTG ATG TC – 3 ‚(23 mer) with a length of 175 bp amplicon DNA and beta lactoglobulin LGB F: 5‘ – TTG TGC TGG ACA GCC ACT AC – 3 ‚(20 mer) LGB R: 5‘ – GCA GCG AGG ACC ACA CAG – 3 ‚(18 mer) with a length of 142 bp DNA amplicon. Then the melting curve analysis of specific heteroduplex DNA with high resolution (HRM) is performed in Light Cycler 480 II Real Time PCR system. HRM is newly applied and optimized for determination of genetic polymorphism of kappa casein and beta-lactoglobulin in cattle. The method determines the genotypes of kappa casein, variants A, B and E and beta-lactoglobulin, variants A and B accurately and simultaneously. |
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