Ligation method employing rtcb
AssigneeAgilent Technologies, Inc.
InventorsGusti Zeiner, Robert A. Ach
Life of this patent
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Abstract
A method of processing an RNA sample is provided. In certain embodiments, the method may comprise: a) obtaining a fragmented RNA sample comprising: i. RNA fragments of long RNA molecules; and ii. unfragmented short RNA; and b) contacting said fragmented RNA sample with a first adaptor in the presence of a RtcB ligase, thereby producing a ligated RNA sample comprising adaptor-ligated fragments of long RNA. A kit for performing the method is also provided.
Claims20
A method of processing a heterogeneous RNA sample comprising:
- a) obtaining a fragmented RNA sample comprising:
- i. RNA fragments of long RNA molecules;
- and
- ii. unfragmented short RNA molecules;
- and
- b) contacting said fragmented RNA sample with a first adaptor in the presence of a RtcB ligase, thereby producing a ligated RNA sample comprising adaptor-ligated fragments of long RNA and unligated short RNAs.
The method of 6, wherein method further comprises contacting said ligated short and long RNA fragments with a third adaptor comprising a 5′-phosphate and a 3′-phosphate which may be in 2′OH,3′P or 2′,3′-cyclic phosphate form, in the presence of a RtcB ligase, thereby producing a ligated RNA sample comprising adaptor-flanked long and short RNAs comprising a 5′-phosphate-end and a blocked 3′-end.
A kit comprising:
- a thermostable RtcB ligase;
- a phosphatase that dephosphorylates the 5′-phosphate of an nucleic acid;
- and
- an adaptor that comprises a 5′-OH and a blocked 3 end.
Dependencies are inferred from the claim text.