Degradable adaptors for background reduction
AssigneeRubicon Genomics, Inc.
InventorsEmmanuel Kamberov, John Langmore, Tim Tesmer
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Abstract
The present disclosure provides systems, processes, articles of manufacture, and compositions that relate to the use of degradable adaptors for background reduction in various nucleic acid manipulations. In particular, adaptors are provided that can be degraded to an extent that the degradation products are incapable or are substantially incapable from participating in subsequent reactions, such as ligation, primer extension, amplification, and sequencing reactions.
Claims43
A method for processing a nucleic acid having at least one cleavable base, comprising:
- (a) creating an abasic site at the at least one cleavable base;
- (b) creating a nick in the backbone of the nucleic acid at the abasic site;
- and
- (c) removing at least one nucleotide adjacent to the nick.
A method for preparing a nucleic acid molecule, comprising:
- (a) providing a double stranded nucleic acid molecule;
- (b) ligating a 3′ end of degradable adaptor comprising at least one cleavable base to a 5′ end of the double stranded nucleic acid molecule to produce an oligonucleotide-attached nucleic acid molecule;
- (c) creating an abasic site at the at least one cleavable base;
- (d) creating a nick at the abasic site;
- and
- (e) removing at least one nucleotide adjacent to the nick.
A kit comprising:
- (a) a nucleic acid comprising at least one cleavable base;
- (b) a uracil-DNA glycosylase;
- (c) an apurinic/apyrimidinic endonuclease;
- and
- (d) an exonuclease.
Dependencies are inferred from the claim text.