NOVEL GELS FOR RAPID RESOLUTION OF HIGH MW DNA

基因组DNA图谱需要分离非常大的DNA 分子（ca. 0.1至3000兆碱基对（Mbp））。 而较小的DNA 分子（0.1-20个内切酶对（kbp））被有效且快速地 通过常规的恒定场凝胶电泳（CFGE）解析， 已实现了0.1至10 Mbp范围内的DNA大小的分辨率 仅使用琼脂糖脉冲场凝胶电泳（PFGE）的变体 凝胶。 PFGE所需时间较长（1-3周）， 凝胶中的迁移距离导致DNA的一些断裂。 新基因 正在开发的制图技术要求对宇宙空间的分辨率 在30到100 kbp的大小范围内。 这些和进一步的发展， 限制性片段长度多态性（RFLP）和基因定位 分析将需要简单和快速的分辨率的DNA分子在 大小范围为50 kbp或更大。 本提案描述了综合和 使用新的专利凝胶，预计将提供快速， 高效的电泳分辨率这样大的DNA分子。 这些 预计凝胶将扩展CFGE的分辨尺寸范围，至少高达 0.5-1 Mbp，并且可能能够提供PFGE的性能优势， 好.

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.