摘要
从培养的HL-60细胞中提取出高分子量的基因组DNA,以EcoRⅠ部分酶解,蔗糖密度梯度离心得到15～30kb的插入片段。以PEG8000沉淀,CsCl密度梯度离心提纯EMBL4噬菌体DNA,以EcoRⅠ,EamHⅠ双酶解得到左右臂,连接、包装得到9.7×10~5个重组体.随机检测了4个重组体,其插入片段平均为19kb.对于19kb的插入段,特定DNA序列达到99％检出率所需的重组体为7.3×10~5(Pfu).
The genomic library of Human Promyelocytic Leukemia cell line (HL—60) are constructed by ligating the DNA inserting fragment in 15～30 kb range and the EMBL4 arms with EcoRI cohensive ends, and packaging in vitro. 9.7×10~5 Pfu recombinant bacteriaphages are produced. The analysis of 4 recombinant clones selected at random suggests that the average size of inserting fragment iss 19 kb. To achieve a 99% probability of having a given DNA sequence represented in a library of 19 kb fragment of a mammalian genome the number of recombinants needs 7.3×10~5. So, the library satisfies the requirement.
出处
《北京师范大学学报（自然科学版）》
CAS
CSCD
1992年第1期99-102,共4页
Journal of Beijing Normal University(Natural Science)
基金
国家自然科学基金
