US 12,486,539 B2
Discriminating BRAF mutations
Amber C. Donahue, Rancho Santa Margarita, CA (US); and Yen-lin Peng, Mission Viejo, CA (US)
Assigned to Quest Diagnostics Investments LLC, Secaucus, NJ (US)
Filed by Quest Diagnostics Investments LLC, Secaucus, NJ (US)
Filed on Sep. 20, 2019, as Appl. No. 16/577,510.
Application 16/577,510 is a continuation of application No. 15/120,345, granted, now 10,422,008, previously published as PCT/US2015/016759, filed on Feb. 20, 2015.
Claims priority of provisional application 61/943,577, filed on Feb. 24, 2014.
Prior Publication US 2020/0115760 A1, Apr. 16, 2020
Int. Cl. C12Q 1/6886 (2018.01)
CPC C12Q 1/6886 (2013.01) [C12Q 2600/106 (2013.01); C12Q 2600/118 (2013.01); C12Q 2600/156 (2013.01)] 6 Claims
OG exemplary drawing
 
1. A kit for distinguishing a V600K mutation from a V600E mutation in the BRAF gene in a sample DNA from an individual, said kit comprising:
(a) a forward primer and a reverse primer and no more than two primers, wherein the forward primer and the reverse primer selectively generate an amplicon when a thymidine (T) to adenosine (A) mutation at position 1799 of the BRAF gene is present, wherein the primers are configured to generate an amplicon comprising a V600E (1799T>A) mutation and an amplicon comprising a V600K (1798-1799GT>AA) mutation in the BRAF gene, and
(b) a restriction enzyme for distinguishing a V600K mutation from a V600E mutation in the BRAF gene, wherein the restriction enzyme digests the amplicon comprising the BRAF V600E mutation (1799T>A) and does not digest the amplicon comprising the BRAF V600K mutation (1798_1799GT>AA),
wherein the forward and/or reverse primer are labeled with a detectable moiety.