MATE PAIR LIBRARY CONSTRUCTION
2 Assignments
0 Petitions
Accused Products
Abstract
The present invention provides a novel method for ligating an adapter to a target polynucleotide and methods of generating a library of mate-pair polynucleotide constructs that employ such a ligation method. Libraries and arrays comprising mate-pair polynucleotide constructs, and methods of sequencing libraries and arrays comprising mate-pair polynucleotide constructs, are also provided.
-
Citations
24 Claims
-
1-7. -7. (canceled)
-
8. A method of making a mate pair polynucleotide library comprising:
-
providing a plurality of double-stranded target polynucleotides; producing circular constructs, each comprising a target polynucleotide, a first adapter, and a nick or gap in the first adapter; performing controlled nick translation to produce nick translation products, each comprising the target polynucleotide, the first adapter, and a nick or gap a first selected distance within the target polynucleotide; performing 3′
branch ligation to ligate a 3′
branch adapter to each nick translation product at the nick or gap to produce gap ligation products;performing controlled primer extension to produce primer extension products by hybridizing a primer to the 3′
branch adapter of the gap ligation products and extending the primer a second selected distance within the target polynucleotides; andadding a 5′
adapter to a 5′
end of the primer extension products to produce a mate pair library, each member of the library comprising;
the 5′
adapter, a first end portion of a target polynucleotide, the first adapter, a second end portion of the target polynucleotide, and the 3′
branch adapter. - View Dependent Claims (9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
-
Specification