METHODS FOR PRODUCING A PAIRED TAG FROM A NUCLEIC ACID SEQUENCE AND METHODS OF USE THEREOF

摘要

Methods for producing a paired tag from a nucleic acid sequence are provided in which the paired tag comprises the 5′ end tag and 3′ end tag of the nucleic acid sequence. In one embodiment, the nucleic acid sequence comprises two restriction endonuclease recognition sites specific for a restriction endonuclease that cleaves the nucleic acid sequence distally to the restriction endonuclease recognition sites. In another embodiment, the nucleic acid sequence further comprises restriction endonuclease recognition sites specific for a rare cutting restriction endonuclease. Methods of using paired tags are also provided. In one embodiment, paired tags are used to characterize a nucleic acid sequence. In a particular embodiment, the nucleic acid sequence is a genome. In one embodiment, the characterization of a nucleic acid sequence is karyotyping. Alternatively, in another embodiment, the characterization of a nucleic acid sequence is mapping of the sequence. In a further embodiment, a method is provided for identifying nucleic acid sequences that encode at least two interacting proteins.

主权项

1. A method for producing a paired tag from a first nucleic acid sequence fragment, without cloning, comprising the steps of:
a) joining the 5′ and 3′ ends of a first nucleic acid sequence fragment via a first linker such that the first linker is located between the 5′ end and the 3′ end of the first nucleic acid sequence fragment in a circular nucleic acid molecule; b) cleaving the circular nucleic acid molecule, thereby producing a second nucleic acid fragment comprising paired tag wherein a 5′end tag of the first nucleic acid sequence fragment is joined to a 3′ end tag of the first nucleic acid sequence fragment via the first linker.