Microfluidic sequencing by hybridization with quantum-dot barcode labels

拟议项目的目标是：i)使用基于液滴的微流控技术生成微型荧光标记库；ii)使用这些标记对DNA链库进行编码，然后用于微流控芯片中的杂交测序。一组不同的荧光量子点将被封装到微流控装置中产生的聚合物微球中。利用多个不同的可区分的强度级别和发射波长，可以产生大量的唯一标签。这些类似条形码的标签将用于杂交的DNA测序，以编码一个DNA寡聚体探针链的大型文库。DNA探针链及其条形码标签包含在油包水乳状液的同一液滴中，并与包含模板的第二液滴合并。杂交事件和探针链通过其条形码标记的身份可以通过荧光检测读出。基于液滴的微流控测序是一种新的方法，有望实现高并行度和最小试剂用量，从而实现快速和成本效益。

Goal of the proposed project is i) to use droplet-based microfluidics to generate a library of micro-sized, fluorescent labels and ii) to use these labels to encode a library of DNA strands that can then be used for sequencing by hybridization in a microfluidic chip. A set of different fluorescent quantum dots will be encapsulated into polymeric microbeads produced in a microfluidic device. With a number of different distinguishable intensity levels and emission wavelengths, a large number of unique labels can be generated. These barcode-like labels will be used for DNA sequencing by hybridization, to encode a large library of DNA oligomer probe strands. The DNA probe strand its barcode label are contained in the same droplet of a water-in-oil emulsion and merged with a second droplet containing the template. The hybridization event and the identity of the probe strand via its barcode label can be read out by fluorescence detection. Microfluidic sequencing based on droplets is a new approach expected to allow for high parallelization and minimal reagent volumes, resulting in high speed and cost efficiency.