Patent ID: 9708657
Date: 2017-07-18
CPC Classifications: C12Q,G16B,G16H,G16Z,Y02A

Claim:
1. A method of generating clonotype profiles from a plurality of immune cell receptor chains, the method comprising the steps of: (a) combining in a reaction mixture under primer extension conditions a first set of primers with a sample of recombined nucleic acids from B-cells and/or T-cells and/or cell-free DNA, wherein each primer of the first set comprises a receptor-specific portion, a 5′-non-complementary end containing a first primer binding site and a sequence tag disposed between the receptor-specific portion and the first primer binding site, wherein the receptor-specific portion anneals to a different recombined nucleic acid at a first predetermined location and is extended to form a first extension product; (b) adding to the reaction mixture under primer extension conditions a second set of primers, wherein each primer of the second set has a receptor-specific portion, wherein the receptor-specific portion anneals to the first extension product at a second predetermined location, and wherein each primer of the second set is extended to form a second extension product, wherein each second extension product comprises a first primer binding site, a sequence tag, and recombined nucleic acid encoding a portion of a T cell receptor chain or a B cell receptor chain; (c) performing a polymerase chain reaction in the reaction mixture to form an amplicon, the polymerase chain reaction using forward primers specific for the first primer binding site and reverse primers specific for the second set of primers; and (d) sequencing the amplicon to form a clonotype profile of a plurality of T cell receptor chains and/or B cell receptor chains.