Use of transposase and Y adapters to fragment and tag DNA
AssigneeAgilent Technologies, Inc.
InventorsRobert A. Ach, Nicholas M. Sampas, Brian Jon Peter
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Abstract
Described herein is an adapter comprising a population of first oligonucleotides, a second oligonucleotide and a third oligonucleotide, wherein the first oligonucleotides, the second oligonucleotide and the third oligonucleotide are hybridized together to produce a complex that comprises: (i) a first end comprising a transposase recognition sequence, (ii) a central single-stranded region of variable sequence and (iii) a second end comprising sequences that are non-complementary. A method, as well as a kit for practicing the method, are also provided.
Claims13
A method for tagmenting a sample, comprising:
- contacting a sample comprising double-stranded DNA with a transposase loaded with adapters;
- wherein the adapters comprise a population of first oligonucleotides, a second oligonucleotide and a third oligonucleotide, wherein the first oligonucleotides, the second oligonucleotide and the third oligonucleotide are hybridized together to produce a complex that comprises:
- (i) a first end comprising a transposase recognition sequence, (ii) a central single-stranded region of variable sequence and (iii) a second end comprising sequences that are non-complementary;
- and
- filling in and sealing the central single-stranded region of the adaptor using a polymerase and ligase, thereby producing a population of DNA fragments that are tagged at both ends by a Y adaptor each comprising the variable sequence of a first oligonucleotide on both strands.
Dependencies are inferred from the claim text.