NOVEL GELS FOR RAPID RESOLUTION OF HIGH MW DNA

绘制基因组DNA图谱将需要分离非常大的DNA 分子(约0.1至3000兆碱基对(MBP))。而较小的DNA 分子(0.1-20千碱基对(KBP))是高效和快速的 通过常规的恒场凝胶电泳法(CFGE)分离， 已经实现了0.1到10 MBP范围内的DNA大小的分辨率 仅在琼脂糖凝胶中使用脉冲场凝胶电泳(PFGE)的变体 凝胶。PFGE所需的较长时间跨度(1-3周)和较长总时间 凝胶中的迁移距离会导致DNA的一些分解。新基因 正在开发需要宇宙星分辨率的测绘技术 在30到100 kBP的尺寸范围内。这些和进一步的发展在 限制性片段长度多态性(RFLP)与粗略基因定位 分析将需要简单而快速地分辨DNA分子在 尺寸范围为50KBP或更大。这项建议描述了合成和 使用新的专有凝胶，预计将提供快速和 如此大的DNA分子的高效电泳法分辨率。这些 凝胶有望将CFGE的分辨率范围扩大到至少 0.5-1 Mbps，并可能使用PFGE AS提供性能优势 井。

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.