Methods of Library Preparation
AssigneeIllumina, Inc.
InventorsAllison Yunghans, Angelica Marie Barr Schalembier, Kayla Busby, Stephen M. Gross
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Abstract
Disclosed herein is a modified transposon end sequence comprising a mosaic end sequence, wherein the mosaic end sequence comprises one or more mutation as compared to a wild-type mosaic end sequence, wherein the mutation comprises a substitution with a uracil, an inosine, a ribose, an 8-oxoguanine, a thymine glycol, a modified purine, or a modified pyrimidine. Also disclosed are transposome complexes comprising these modified transposon end sequences and methods of library preparation using these modified transposon end sequences.
Claims20
A modified transposon end sequence comprising a mosaic end sequence, wherein the mosaic end sequence comprises one or more mutations as compared to a wild-type mosaic end sequence, wherein the mutation comprises a substitution with
- a. a uracil;
- b an inosine;
- c. a ribose;
- d. an 8-oxoguanine;
- e. a thymine glycol;
- f. a modified purine;
- or
- g. a modified pyrimidine.
A transposome complex comprising:
- a. a transposase;
- b. a first transposon comprising a modified transposon end sequence comprising a uracil, an inosine, a ribose, an 8-oxoguanine, a thymine glycol, a modified purine, and/or a modified pyrimidine;
- and
- c. a second transposon comprising a second transposon end sequence complementary to at least a portion of the first transposon end sequence.
A method of preparing double-stranded nucleic acid fragments comprising adapters comprising:
- a. combining a sample comprising nucleic acid with transposome complexes comprising:
- i. a transposase;
- ii a first transposon comprising a modified transposon end sequence comprising a uracil, an inosine, a ribose, an 8-oxoguanine, a thymine glycol, a modified purine, and/or a modified pyrimidine;
- and
- iii. a second transposon comprising a second transposon end sequence complementary to at least a portion of the first transposon end sequence;
- b. preparing nucleic acid fragments;
- c. combining the sample with (1) an endonuclease or (2) a combination of a DNA glycosylase and heat, basic conditions, or an endonuclease/lyase that recognizes abasic sites and cleaving the first transposon end at the uracil, inosine, ribose, 8-oxoguanine, thymine glycol, modified purine, and/or modified pyrimidine within the mosaic end sequence to remove all or part of the first transposon end from the nucleic acid fragments;
- and
- d. ligating an adapter onto the 5′ and/or 3′ ends of the nucleic acid fragments.
Dependencies are inferred from the claim text.