Physical mapping of complex genomes
A method for the simultaneous identification of overlapping cosmid clones among multiple cosmid clones and the use of the method for mapping complex genomes are provided. A library of cosmid clones that contains the DNA to be mapped is constructed and arranged in a manner such that individual clones can be identified and replicas of the arranged clones prepared. In preferred embodiments, the clones are arranged in a two dimensional matrix. In such embodiments, the cosmid clones in a row are pooled, mixed probes complementary to the ends of the DNA inserts in the pooled clones are synthesized, hybridized to a first replica of the library. Hybridizing clones, which include the pooled row, are identified. A second portion of clones is prepared by pooling cosmid clones that correspond to a column in the matrix. The second pool thereby includes one clone from the first portion pooled clones. This common clone is located on the replica at the intersection of the column and row. Mixed probes complementary to the ends of the DNA inserts in the second pooled portion of clones are prepared and hybridized to a second replica of the library. The hybridization pattern on the first and second replicas of the library are compared and cross-hybridizing clones, other than the clones in the pooled column and row, that hybridize to identical clones in the first and second replicas are identified. These clones necessarily include DNA inserts that overlap with the DNA insert in the common clone located at the intersection of the pooled row and pooled column. The DNA in the entire library may be mapped by pooling the clones in each of the rows and columns of the matrix, preparing mixed end-specific probes and hybridizing the probes from each row or column to a replica of the library. Since all clones in the library are located at the intersection of a column and a row, the overlapping clones for all clones in the library may be identified and a physical map constructed.
- DOE Contract Number:
- FG03-88ER60694
- Assignee:
- Salk Inst. for Biological Studies, San Diego, CA (United States)
- Patent Number(s):
- US 5219726; A
- Application Number:
- CNN: R01HD18012; PPN: US 7-360254
- OSTI ID:
- 6221430
- Resource Relation:
- Patent File Date: 2 Jun 1989
- Country of Publication:
- United States
- Language:
- English
Similar Records
Cloning human chromosome 9-specific cDNAs by an efficient selection strategy using somatic hybrids
Sequence Ready Characterization of the Pericentromeric Region of 19p12