Klinman D M, Cohen D I
Anal Biochem. 1987 Feb 15;161(1):85-8. doi: 10.1016/0003-2697(87)90655-5.
A technique for the long term storage of primary cDNA libraries in a form such that relevant DNA sequences can be readily identified and retrieved is described. cDNA libraries produced using the lambda gt 10 cloning vector were plated out on host bacteria in 0.7% top agarose supplemented with 30% glycerol. Nitrocellulose lifts of these libraries were made and stored. These lifts could be screened at a later time to permit identification of bacteriophage plaques containing specific cDNA inserts. The plated libraries were then transferred to a -70 degrees C freezer. The combination of freezing and glycerol treatment allowed the bacteriophage in these primary cDNA libraries to remain viable for significantly longer than 1 year.
本文描述了一种用于长期保存原始cDNA文库的技术,该文库形式能使相关DNA序列易于识别和检索。使用λgt 10克隆载体产生的cDNA文库铺板于含有30%甘油的0.7%顶层琼脂糖中的宿主细菌上。制备这些文库的硝酸纤维素滤膜并保存。这些滤膜可在以后进行筛选,以鉴定含有特定cDNA插入片段的噬菌体噬菌斑。然后将铺板的文库转移至-70℃冰箱。冷冻和甘油处理相结合,使这些原始cDNA文库中的噬菌体存活时间显著超过1年。