US 10,889,850 B2
Methods for improved isolation of genomic DNA templates for allele detection
Connie Chao-Shern, Menlo Park, CA (US)
Assigned to Avellino Lab USA, Inc., Menlo Park, CA (US)
Filed by Avellino Lab USA, Inc., Menlo Park, CA (US)
Filed on Jan. 2, 2018, as Appl. No. 15/860,051.
Application 15/860,051 is a continuation in part of application No. 14/788,572, filed on Jun. 30, 2015, granted, now 9,856,516.
Application 14/788,572 is a continuation of application No. PCT/US2014/029466, filed on Mar. 14, 2014.
Claims priority of provisional application 61/852,357, filed on Mar. 15, 2013.
Claims priority of provisional application 61/852,358, filed on Mar. 15, 2013.
Prior Publication US 2018/0230514 A1, Aug. 16, 2018
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C07H 21/00 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6883 (2018.01); C12N 15/10 (2006.01)
CPC C12Q 1/6806 (2013.01) [C12N 15/1003 (2013.01); C12Q 1/6883 (2013.01); C12Q 2600/156 (2013.01)] 21 Claims
 
1. A method for preparing genomic DNA (gDNA) samples, the method comprising:
(A) providing a sample of buccal epithelial cells from a subject, the buccal epithelial cells adhered to at least a part of a substrate;
(B) agitating, for no more than 120 seconds, said at least a part of the substrate with the adhered epithelial cells in a first lysis solution capable of lysing buccal cells adhered to the substrate;
(C) removing the substrate from the first lysis solution;
(D) performing an extraction incubation of gDNA in the first lysis solution by incubating the first lysis solution at a temperature between 35° C. and 55° C. for an incubation time between 15 minutes and 120 minutes after removing (C) the substrate from the first lysis solution to prepare a first gDNA sample;
(E) agitating said at least a part of the substrate, after removing the substrate from the first lysis solution, in a second lysis solution capable of lysing buccal cells adhered to the substrate;
(F) removing the substrate from the second lysis solution; and
(G) performing an extraction incubation of second gDNA in the second lysis solution by incubating the second lysis solution at a temperature between 35° C. and 55° C. for an incubation time between 15 minutes and 120 minutes after removing (F) the substrate from the second lysis solution to prepare a second gDNA sample.