US 12,280,350 B2
Compositions and methods for the amplification of nucleic acids
Jian-Bing Fan, San Diego, CA (US); Kevin L. Gunderson, Encinitas, CA (US); Frank J. Steemers, Encinitas, CA (US); Dmitry K. Pokholok, San Marcos, CA (US); and Fiona Kaper, San Diego, CA (US)
Assigned to Illumina, Inc., San Diego, CA (US)
Filed by Illumina, Inc., San Diego, CA (US)
Filed on Nov. 16, 2020, as Appl. No. 17/098,840.
Application 17/098,840 is a continuation of application No. 14/416,563, granted, now 10,870,099, previously published as PCT/US2013/027017, filed on Feb. 21, 2013.
Claims priority of provisional application 61/676,153, filed on Jul. 26, 2012.
Prior Publication US 2021/0129107 A1, May 6, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. B01J 19/00 (2006.01); C12Q 1/6844 (2018.01)
CPC B01J 19/0046 (2013.01) [C12Q 1/6844 (2013.01); B01J 2219/00585 (2013.01); B01J 2219/00722 (2013.01)] 20 Claims
 
1. A method of preparing a nucleic acid library from a nucleic acid sample, the method comprising:
(a) providing a set of amplification primers comprising:
a plurality of random primers, wherein each random primer comprises a random 3′ portion, a constant 5′ priming portion and at least one nucleotide analogue comprising a non-natural base located between the random 3′ portion and the constant 5′ priming portion;
a plurality of locus specific primers configured to amplify a plurality of predetermined genomic regions; and
a plurality of blocked primers;
wherein each plurality of random primers and locus specific primers in the set of amplification primers is a mixture of different primers;
(b) hybridizing the set of amplification primers to the nucleic acid sample and extending the hybridized amplification primers to obtain extended polynucleotides, wherein the extending is performed in the absence of a nucleotide capable of hybridizing to the at least one nucleotide analogue;
(c) circularizing the extended polynucleotides to obtain circular polynucleotides; and
(d) amplifying the circular polynucleotides in the presence of the nucleotide capable of hybridizing to the at least one nucleotide analogue, thereby preparing the nucleic acid library.