Method and compositions for ordering restriction fragments
First Claim
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1. 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.
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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.
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Citations
5 Claims
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1. 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.- View Dependent Claims (2, 3, 4, 5)
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Specification