基因组YAC文库构建方法的几点改进

Some Improved Methods for the Construction of Genomic YAC Library.

以ｐＪＳ９７、ｐＪＳ９８为载体，对中国人基因组ＹＡＣ文库的构建进行了尝试建立了一种对小片段ＤＮＡ“原位电泳筛除”的方案，采用多胺溶液处理以减少大片段ＤＮＡ的降解，使整个建库工作更加简便、有效．此外，改进了转化子的挑取和保存的方法，既简化了操作，又减少了杂菌污染和交叉污染．这些改进的方案。

A Chinese human genomic YAC (yeast artificial chromosome) library is being constructed with pJS97 and pJS98 as cloning vectors. An easier and more efficient protocol has been made successfully to protect the high molecular weight DNA from being degraded byusing polyamines solution and to remove smaller DNA by setting up a new method called 'in situ electrophoretic size-fractionation'.Furthermore, the glycerol at the final concentration of 20% is introduced directly into the medium used to grow the transformants, which would both simplify the operation and reduce the chance of contamination with other microorganisms or cross-contamination of YAC clones. Such improved methods can be used in the construction and application of other genomic YAC libraries.