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US 12,686,863 B2Granted
Methods and compositions for rapid nucleic library preparation
AssigneeTwist Bioscience Corporation
InventorKeith Brown
Life of this patent
Estimating the term…
Abstract
Rapid nucleic acid libraries, methods of generation, kits, and compositions relating to library synthesis, including reagents, intermediaries and final products are disclosed herein. The disclosure enables rapid synthesis of libraries that allow independent verification of sequence information and rapid identification of sequence information with template of origin.
Claims19
A kit for generating a library from a target nucleic acid sample, the kit comprising:
- an extension mixture comprising dNTPs and ddNTPs;
- a first primer library comprising a first plurality of random primers configured to generate a population of non-identical sequences each comprising a subset of a target nucleic acid sample and at least one mismatch between a random primer of the first plurality of random primers and the target nucleic acid sequence;
- and
- a second primer library comprising a second plurality of random primers configured to extend the population of non-identical sequences to obtain double-stranded sequences.
1 more element
A kit for generating a library from a target nucleic acid sample, the kit comprising:
- an extension mixture comprising dNTPs and ddNTPs, wherein a molar ratio of dNTPs:ddNTPs is at least about 49 dNTPs to 1 ddNTP;
- and
- a primer library comprising a plurality of random primers, wherein the primer library is configured to generate a population of non-identical sequences each comprising a subset of a target nucleic acid sample and at least one mismatch between a random primer of the plurality of random primers and the target nucleic acid sample.
A kit for generating a library from a target nucleic acid sample, the kit comprising:
- an extension mixture comprising dNTPs and ddNTPs;
- and
- a primer library comprising a plurality of random primers, wherein a GC content of the primer library is biased to match a relative abundance of GC content in a target nucleic acid sequence, and wherein the primer library is configured to generate a population of non-identical sequences each comprising a subset of the target nucleic acid sample and at least one mismatch between a random primer of the plurality of random primers and the target nucleic acid sequence.
Dependencies are inferred from the claim text.