Method of Making a Paired Tag Library for Nucleic Acid Sequencing

• US 20150315639A1
• Filed: 07/14/2015
• Published: 11/05/2015
• Est. Priority Date: 01/09/2008
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Li, Bin, Xi, Lei, Ranade, Swati, Wang, Yangzhou
• Granted Patent: US 9,657,291 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2521/325 Single stranded exonuclease
• CPC Class Codes: C12Q 2525/191 incorporating an adaptor
• CPC Class Codes: C12Q 2525/307 Circular oligonucleotides

Abstract

The present disclosure relates to methods and compositions for making paired tags and paired tag libraries.

Claims

1-91. -91. (canceled)

92. A plurality of circular nucleic acid molecules, the individual circular molecules in the plurality comprising:

(a) a universal sequence region common to the plurality of circular nucleic acid molecules and having two nicks; and

(b) a target sequence region that differs between different molecules of the plurality of circular molecules.

93. The molecules of claim 92, wherein the universal sequence region comprises a double-stranded nucleic acid.

94. The molecules of claim 92, wherein the universal sequence further comprises a binding moiety.

95. The molecule of claim 94, wherein the binding moiety comprises biotin.

96. The molecules of claim 94, further comprising a purifying moiety.

97. The molecules of claim 96 wherein the purifying moiety comprises streptavidin.

98. The molecule of claim 92, further comprising a nick translating enzyme.

99. The molecule of claim 98, wherein the nick translating enzyme comprises DNA polymerase I.

100. A plurality of circular nucleic acid molecules, the individual circular molecules in the plurality comprising:

(a) a universal sequence region common to the plurality of circular nucleic acid molecules, the universal sequence region having a first end and a second end;

(b) a target sequence region that differs between different molecules of the plurality of circular molecules, the target sequence region having a first end and a second end;

(c) a first nick between the first end of the universal sequence region and the first end of the target sequence region; and

(d) a second nick between the second end of the universal sequence region and the second end of the target sequence region.