Patent ID: 9624533
Date: 2017-04-18
CPC Classifications: C12Q

Claim:
1. A method for multiplex ligation-dependent probe amplification comprising: (a) providing a sample tissue to query at least 100 different target nucleic acids; (b) providing a plurality of different probe sets for each of the at least 100 different target nucleic acids, wherein each probe set comprises: (c) hybridizing the plurality of different probe sets to the at least 100 different target sequences to form at least 100 different hybridization complexes; (d) ligating, in a single vessel or tube, the at least 100 different hybridization complexes to form at least 100 different ligated probes; (e) amplifying the at least 100 different ligated probes to form at least 100 different amplicons, wherein (i) the amplifying step is carried out with a first universal primer comprising a region complementary to the first adapter sequence and a second universal primer comprising a region complementary to the second adapter sequence, and (ii) the amplifying of the at least 100 different ligated probes is carried out in the single vessel or tube; and (f) detecting the at least 100 different amplicons in a detection system, independently of the length, by sequencing each of the at least 100 different amplicons wherein said at least 100 different ligated probes of step (d) are directly amplified in step (e) without purification.