cDNA library for nucleic acid sequencing
First Claim
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1. A method for preparing a library of DNA copies of a plurality of RNA molecules, comprising:
- (a) fragmenting the plurality of RNA molecules to produce RNA fragments having a 5′
end and a 3′
end;
(b) producing RNA constructs by adding a 5′
-RNA oligonucleotide adaptor segment so that it becomes ligated to the 5′
end of the RNA fragments and adding a 3′
-RNA oligonucleotide adaptor segment so that it becomes ligated to the 3′
end of the RNA fragments;
(c) preparing double-stranded DNA copies of the RNA constructs; and
(d) producing a library of circularized double-stranded DNA copies of said RNA fragments, wherein the circularized DNA copies comprise an adaptor that contains a complete DNA copy of the entire 5′
-RNA oligonucleotide adaptor segment joined to the entire 3′
-RNA oligonucleotide adaptor segment.
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Abstract
The present invention is directed to compositions and methods for nucleic acid identification and detection. Compositions and methods of the present invention include extracting and fragmenting target nucleic acids from a sample, using the fragmented target nucleic acids to produce target nucleic acid templates and subjecting those target nucleic acid templates to amplification methods to form nucleic acid nanoballs. The invention also includes methods of detecting and identifying sequences using various sequencing applications, including sequencing by ligation methods.
34 Citations
8 Claims
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1. A method for preparing a library of DNA copies of a plurality of RNA molecules, comprising:
-
(a) fragmenting the plurality of RNA molecules to produce RNA fragments having a 5′
end and a 3′
end;(b) producing RNA constructs by adding a 5′
-RNA oligonucleotide adaptor segment so that it becomes ligated to the 5′
end of the RNA fragments and adding a 3′
-RNA oligonucleotide adaptor segment so that it becomes ligated to the 3′
end of the RNA fragments;(c) preparing double-stranded DNA copies of the RNA constructs; and (d) producing a library of circularized double-stranded DNA copies of said RNA fragments, wherein the circularized DNA copies comprise an adaptor that contains a complete DNA copy of the entire 5′
-RNA oligonucleotide adaptor segment joined to the entire 3′
-RNA oligonucleotide adaptor segment. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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Current AssigneeComplete Genomics Incorporated (BGI Genomics Co., Ltd.)
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Original AssigneeComplete Genomics Incorporated (BGI Genomics Co., Ltd.)
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InventorsDrmanac, Radoje, Dahl, Fredrik, Hurowitz, Evan, Dahl, Fredrie
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Primary Examiner(s)BOESEN, CHRISTIAN C
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Application NumberUS12/956,802Publication NumberTime in Patent Office1,617 DaysField of SearchNoneUS Class Current506/26CPC Class CodesB01J 2219/00621 by physical means, e.g. tre...B01J 2219/00637 by coating it with another ...B01J 2219/00659 Two-dimensional arraysB01J 2219/00722 NucleotidesC12N 15/10 Processes for the isolation...C12Q 1/6853 using modified primers or t...C12Q 1/6869 Methods for sequencingC40B 20/00 Methods specially adapted f...
