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US 12,180,468 B2Granted
Transposase compositions for reduction of insertion bias
AssigneeIllumina, Inc.
InventorsChristian Gloeckner, Amirali Kia, Molly He, Trina Faye Osothprarop, Frank J. Steemers, Kevin L. Gunderson, Sasan Amini, Jerome Jendrisak
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Abstract
Presented herein are methods and compositions for tagmentation of nucleic acids. The methods are useful for generating tagged DNA fragments that are qualitatively and quantitatively representative of the target nucleic acids in the sample from which they are generated.
Claims17
A method for preparing a nucleic acid library, comprising:
- (a) contacting a target nucleic acid with a first transposome in a reaction buffer comprising manganese cations, thereby generating a first plurality of tagmented nucleic acids, wherein the first transposome comprises a Tn5 transposase;
- (b) contacting the first plurality of tagmented nucleic acids with a second transposome to generate a second plurality of tagmented nucleic acids, wherein the second transposome comprises a Mos-1 transposase;
- and
- (c) amplifying the second plurality of tagmented nucleic acids to generate a nucleic acid library having an increased representation of different sequences of the target nucleic acid compared to a nucleic acid library prepared by a single tagmentation of the target nucleic acid.
1 more element
A method for preparing a nucleic acid library, comprising:
- (a) generating a first plurality of tagmented nucleic acids by contacting a target nucleic acid with a first transposome in the presence of manganese cations, wherein the first transposome comprises a Tn5 transposase;
- (b) generating a second plurality of tagmented nucleic acids by contacting the first plurality of tagmented nucleic acids with a second transposome, wherein the second transposome comprises a Mos-1 transposase;
- and
- (c) amplifying the second plurality of tagmented nucleic acids to generate a nucleic acid library.
1 more element
A method for preparing a nucleic acid library, comprising:
- (a) contacting a target nucleic acid with a first transposome in a reaction buffer to generate a first plurality of tagmented nucleic acids, wherein the first transposome comprises a Tn5 transposase;
- (b) adjusting a salt concentration of the reaction buffer by addition of NaCl;
- (c) contacting the first plurality of tagmented nucleic acids with a second transposome to generate a second plurality of tagmented nucleic acids, wherein the second transposome comprises a Mos-1 transposase;
- and
- (d) amplifying the second plurality of tagmented nucleic acids to generate a nucleic acid library having an increased representation of different sequences of the target nucleic acid compared to a nucleic acid library prepared by a single tagmentation of the target nucleic acid.
2 more elements
Dependencies are inferred from the claim text.