NOVEL GELS FOR RAPID RESOLUTION OF HIGH MW DNA

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Mapping of genomic DNA will require the separation of very large DNA molecules (ca. 0.1 to 3000 Megabase-pairs (Mbp)). While smaller DNA molecules (0.1-20 kilobase-pairs (kbp)) are efficiently and rapidly resolved by the conventional Constant Field Gel Electrophoresis (CFGE), the resolution of DNA sizes in the range of 0.1 to 10 Mbp has been achieved only using variants of pulsed-field gel electrophoresis (PFGE) in agarose gels. The long time span (1-3 weeks) needed for PFGE and the long total migration distance in the gel result in some breakdown of DNA. New gene mapping techniques are being developed which require resolution of cosmids in the size range of 30 to 100 kbp. These and further developments in restriction fragment length polymorphism (RFLP) and coarse gene mapping analyses will require simple and rapid resolution of DNA molecules in the size range of 50 kbp and larger. This proposal describes the synthesis and use of new proprietary gels which are expected to provide rapid and efficient electrophoretic resolution of such large DNA molecules. These gels are expected to extend the resolving size range of CFGE at least upto 0.5-1 Mbp and may be able to provide performance advantages with PFGE as well.