C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
195/1.235, 150/8
C12Q 1/68 (2006.01) C07H 21/04 (2006.01)
Patent
CA 2030967
Abstract: In overlapping chromosome fragments to physical map chromosomes it is most desirable to have a single method which combines, simplicity, precision, speed and reproducibility,and produces long contiguous stretches of ordered DNA. Until now there was no method available that meets these criteria. In this invention a, "3rd generation mapping method", which uses a strategy based on a modular contig building procedure and a "vector-cloning site- implant", all the above desirable criteria are met. In this method unique "fragment ends" copied by the polymerase chain reaction (PCR) using two pairs of primers, from uniformily sized fragments produced by complete digestion of chromosomes with selected 6-base blunt cutting restriction enzymes, are used to completely overlap and orientate vectors carrying chromosome fragments. The method is applicable to all types of vectors and all sizes of fragments. Vectors carrying implants are called RSCC vectors. Overlapping is fast, precise, efficient (little or no redundancy) and reproducible. In addition, the ends used to overlap fragments in one library are portable to any other library of the same chromosome, where they can be used to group clones in the library into "contigs members" before they are mapped.
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