METHODS FOR IMPROVED ISOLATION OF GENOMIC DNA TEMPLATES FOR ALLELE DETECTION

摘要

The present disclosure relates to improved methods for the isolation of genomic material and detection of disease related single nucleotide polymorphisms. In some aspects, these methods increase the total recovery of genomic DNA from buccal cell samples by improving cell lysis conditions. In other aspects, these methods allow for the reuse of patient buccal swab samples, reducing the likelihood of having to collect additional patient samples for re-testing. Finally, in some aspects, these methods increase the sensitivity of SNP detection using an improved real-time PCR assay protocol.

主权项

1. A method for preparing a genomic DNA (gDNA) sample for detection of a single nucleotide polymorphism, the method comprising:
(A) providing a sample of buccal epithelial cells from a subject, the buccal epithelial cells adhered to the tip of a substrate; (B) agitating, for no more than about 45 seconds, the tip of the substrate with the adhered epithelial cells in a first lysis solution capable of lysing buccal cells adhered to the substrate for no more than about 45 seconds; (C) removing the substrate from the first lysis solution upon completion of the agitating (B) the tip of the substrate in the first lysis solution; and (D) performing an extraction incubation of gDNA in the first lysis solution by incubating the first lysis solution at a temperature of 45±3° C. for 45±15 minutes after removing (C) the substrate from the first lysis solution.