Method and compositions for ordering restriction fragments
AssigneeLynx Therapeutics, Inc.
InventorStephen C. Macevicz
Life of this patent
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Abstract
The invention provides a method for constructing a high resolution physical map of a polynucleotide. In accordance with the invention, nucleotide sequences are determined at the ends of restriction fragments produced by a plurality of digestions with a plurality of combinations of restriction endonucleases so that a pair of nucleotide sequences is obtained for each restriction fragment. A physical map of the polynucleotide is constructed by ordering the pairs of sequences by matching the identical sequences among the pairs.
Claims5
A vector containing a restriction fragment of DNA, the vector having a top strand and a bottom strand, and comprising, in sequence:
- a first type IIs recognition site oriented in the 5′ to 3′ direction of said top strand,
- said restriction fragment, and
- a second type IIs recognition site oriented in the 5′ to 3′ direction of said bottom strand,
- such that a first type IIs restriction endonuclease recognizing the first type IIs recognition site has a cleavage site which is 3′ of the first recognition site in the top strand 5′ to 3′ direction, and is within the restriction fragment,
- and a second type IIs restriction endonuclease recognizing the second type IIs recognition site has a cleavage site which is 3′ of the second recognition site in the bottom strand 5′ to 3′ direction, and is within the restriction fragment.
Dependencies are inferred from the claim text.