McCormick M K, Antonarakis S E, Hieter P
Genetics Group, Los Alamos National Laboratory, NM 87545.
Genet Anal Tech Appl. 1990 Sep;7(5):114-8. doi: 10.1016/0735-0651(90)90016-9.
Yeast artificial chromosome (YAC) cloning of DNA in agarose is an alternative method to cloning from aqueous solutions. It minimizes any shearing that may result from handling of high molecular weight DNA and can be done with nanogram to microgram amounts of material, which facilitates construction of YACs from sources of DNA other than genomic DNA isolated from cells. The average size of the YACs recovered (200-1000 kb) and efficiency of transformation of ligation products (200-1000 cfu/micrograms) are similar to those reported using aqueous protocols. This method has been used to construct chromosome specific YACs, and it should be possible to apply the technique to the construction of chromosome specific libraries using flow sorted chromosomes as source material, and the cloning of restriction fragments isolated by preparative pulsed field gel electrophoresis.
在琼脂糖中进行酵母人工染色体（YAC）DNA克隆是从水溶液中克隆的另一种方法。它能将因处理高分子量DNA而可能产生的任何剪切降至最低，并且可以使用纳克至微克量的材料来完成，这便于从细胞分离的基因组DNA以外的DNA来源构建YAC。回收的YAC的平均大小（200 - 1000 kb）和连接产物的转化效率（200 - 1000 cfu/微克）与使用水性方案报道的相似。该方法已用于构建染色体特异性YAC，并且应该可以将该技术应用于使用流式分选染色体作为源材料构建染色体特异性文库，以及克隆通过制备性脉冲场凝胶电泳分离的限制性片段。
