大规模EST序列测定中的条件优化

The Condition Optimization of the Large-scale EST Sequencing

EST技术已广泛用于发现新基因和基因表达谱研究。笔者对EST序列测定中的不同反应体系、Big Dye类型、用量、测序产物纯化及不同测序板等因素进行了比较,确定了大规模EST测序方案,同时对文库质量及其他影响因素进行了讨论,经优化后的方案可降低大规模EST测序成本及提高测序成功率。

As an effective tool for the research on gene discovery and gene expression profile,expressed sequence tags ( EST) have been taken into account by many laboratories in the world.Reducing the cost of large-scale EST sequencing and improving the success rate of EST sequencing are particularly important for relevant research.Based on different comparison between such factors as response system,BigDye type,content of BigDye,purification of sequence product and 96-well reaction plate,the relative cheap project of large-scale EST sequencing has been confirmed.The quality of cDNA library and other influence factors have been discussed at the same time.It is significant and useful for reducing the cost and improving the success rate of EST sequencing.