NOVEL MATERIALS FOR LOW CONC ACRYLAMIDE LIKE GELS

我们建议开发基于预聚物的胶凝化学， 与DEL测序和蛋白质凝胶电泳兼容， 凝胶的浓度远低于通常的丙烯酰胺/双凝胶。 增加DEL测序的通量的需要是众所周知的。如果 读出速率可以增加到每个反应超过1200个碱基 （B/rxn），重叠测序和序列组装步骤的减少 可能导致净吞吐率增加几倍。如果 凝胶可以在低至3%T（总浓度）下运行，然后碱 预期给定技术的读出速率几乎加倍。因此 自动测序的读出速率超过1500， 可以获得手动技术。市售预聚物可固化 在3%T下足够强的凝胶，但它们的化学性质与 DNA测序 将评估几种不同的预聚物凝胶化学性质： 能够将凝胶固化为5%T丙烯酰胺/双凝胶， %T;以及它们在测序中获得单碱基DNA分辨率的能力。 这些预聚物凝胶也将用于解决临床 重要的大蛋白质和蛋白质-核酸组装体。 建议的商业应用：成功的项目将产生新的 预聚物凝胶用于DEL测序和用于非常大的蛋白质 电泳潜在市场超过1亿美元。预聚 这项技术将使预涂凝胶能够适当地 设计用于快速、确定、DNA导向检测的临床仪器 用于感染、癌症和遗传标记， 几十亿美元。

We propose to develop prepolymer-based gelling chemistries that are compatible with DEL sequencing and protein gel electrophoresis to make gels at concentrations much lower than the usual acrylamide/bis gels. The need for increasing throughput of DEL sequencing is well known. If the readout rate can be increased to more than 1200 bases per reaction (b/rxn), the reduction in overlap sequencing and sequence assembly steps could lead to an increase in the net throughput rate of several-fold. If gels can be run at as low as 3%T (total concentration), then the base readout rate of a given technique is expected to be almost doubled. Thus readout rates of over 1500 in automated sequencing and over 1000b/rxn in manual techniques may be attainable. Commercial prepolymers can set strong enough gels at 3%T, but their chemistries are incompatible with DNA sequencing. Several different prepolymer gel chemistries will be evaluated for: Their ability to set gels as strong as 5%T acrylamide/bis gels at inch lower %T; and their ability to attain single base DNA resolution in sequencing. These prepolymer gels will also be useful for resolving clinically important large proteins, and protein-nucleic acid assemblies. PROPOSED COMMERCIAL APPLICATION: Successful project will result in novel prepolymer gels for DEL sequencing and for very large protein electrophoresis. The potential market is over $100 million. Prepolymer technology will later enable precoated ultrathin gels for appropriately designed clinical instruments for rapid, definitive, DNA-oriented tests for infections, cancers, and genetic markers, This market is expected to be several billion dollars.