Login
欢迎浏览恩派尔生物资料网
我要投稿 请登录 免费注册 安全退出

您现在的位置是: 首页 > 实验方法 > 生物化学

生物化学

Screening Recombinant Libraries by Polymerase Chain Reaction

2025-03-27 生物化学 加入收藏
The complexity of the genome of a particular organism or the relative abundance

The complexity of the genome of a particular organism or the relative abundance of a particular mRNA, within the cell type from which a cDNA library was constructed, affects the ability with which one can isolate a gene or cDNA clone of interest. With respect to genomic libraries, the number of clones needed to be screened to isolate a single-copy sequence is a function of the complexity of the genome and the average size of the cloned fragments in the library (1 ). In the case of cDNA libraries, the frequency of a given clone of interest depends on the abundance of the messenger RNA. Highly abundant messages can represent 10% or more of total mRNA, whereas very rare messages can be as low as one in 106 . In addition, the representation of some sequences in a cDNA library, particularly the 5′ ends of large mRNAs, will be less than expected due to the technical difficulties in converting the mRNA into full-length cDNA copies. In some cases, a particular sequence of interest can be depleted or lost at various steps of screening due to its inefficiency to be replicated relative to other clones in the library.

文章底部广告位

文章评论

加载中~