Methods and compositions for preparing nucleic acid libraries
First Claim
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1. A method for preparing a nucleic acid library from a low quality nucleic acid sample comprising:
- (a) providing a plurality of nucleic acids obtained from the low quality nucleic acid sample, and selectively shearing the plurality of nucleic acids to form sheared nucleic acids by a method comprising;
(i) separating the plurality of nucleic acids into a first fraction of nucleic acids having an average length longer than 550 base pairs, and a second fraction of nucleic acids having an average length shorter than 550 base pairs,(ii) shearing the nucleic acids of the first fraction, and(iii) combining the sheared first fraction and the second fraction to form the sheared nucleic acids;
(b) removing single-stranded nucleic acids from the plurality of sheared nucleic acids; and
(c) repairing the nucleic acids of the plurality of sheared nucleic acids, thereby preparing a nucleic acid library.
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Abstract
Embodiments provided herein relate to methods and compositions for preparing nucleic acid libraries. Some embodiments include preparing libraries from nucleic acids obtained from degraded samples, such as ancient samples and fixed samples.
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22 Claims
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1. A method for preparing a nucleic acid library from a low quality nucleic acid sample comprising:
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(a) providing a plurality of nucleic acids obtained from the low quality nucleic acid sample, and selectively shearing the plurality of nucleic acids to form sheared nucleic acids by a method comprising; (i) separating the plurality of nucleic acids into a first fraction of nucleic acids having an average length longer than 550 base pairs, and a second fraction of nucleic acids having an average length shorter than 550 base pairs, (ii) shearing the nucleic acids of the first fraction, and (iii) combining the sheared first fraction and the second fraction to form the sheared nucleic acids; (b) removing single-stranded nucleic acids from the plurality of sheared nucleic acids; and (c) repairing the nucleic acids of the plurality of sheared nucleic acids, thereby preparing a nucleic acid library. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A method for preparing a nucleic acid library from a low quality nucleic acid sample comprising single stranded DNA, the method comprising:
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(a) providing a plurality of nucleic acids obtained from the low quality nucleic acid sample, the nucleic acids comprising single stranded DNA, and selectively shearing the plurality of nucleic acids to form sheared nucleic acids by a method comprising; (i) separating the plurality of nucleic acids into a first fraction having an average length longer than 550 base pairs, and a second fraction having an average length shorter than 550 base pairs, (ii) shearing the nucleic acids of the first fraction, and (iii) combining the sheared first fraction and the second fraction; (b) hybridizing random primers to the single stranded DNA and extending the primers to form double stranded DNA; and (c) treating the double stranded DNA to remove single strand overhangs to form blunt ended DNA, thereby preparing a nucleic acid library. - View Dependent Claims (18, 19, 20, 21, 22)
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Specification