Multiplex sequencing and cosmid mapping: Progress report
The multiplexing project has progressed on two fronts. Multiplex dideoxy sequencing has been refined and another group is nearly ready to do a bench mark sequence of the maize mitochondrial genome (450kb) to get a real estimate of the speed and efficiency of the method. The 16 overlapping cosmids containing the whole genome have been subcloned onto 3200 clones on 64 multiplexing vectors, which have been divided into 50 fold mixtures. After electro blotting onto 80 membranes, the sequences can be retrieved by probing with P 2-poly-dA-tailed oligos. The turn around time for probing, washing exposing and stripping is about 24 hours. We are building an automated system that has the membranes on the inside of a large rotating drum so that all the steps of the probing cycle are carried out without having to handle the membranes and keeping the hybridization solution to a minimum. The second front is the construction of a florescence array detector that we hope will allow us to replace the P32 probe with a fluorescent probe that will increase speed of detection and add another level of multiplexing.
- Research Organization:
- Utah Univ., Salt Lake City (USA). Dept. of Human Genetics
- DOE Contract Number:
- FG02-88ER60700
- OSTI ID:
- 6219284
- Report Number(s):
- DOE/ER/60700-T1; ON: DE89007710
- Resource Relation:
- Other Information: Portions of this document are illegible in microfiche products
- Country of Publication:
- United States
- Language:
- English
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