全长cDNA文库的构建方法

The Methods of Constructing the Full Length cDNA Library

全长cDNA文库的构建是进行功能基因组研究的一种经济、快速、有效的途径,克服了传统cDNA文库的缺点,节省了基因克隆和功能鉴定的时间,促进了功能基因组的研究进程。目前有6种构建全长cDNA文库的方法,包括Oligo-Capping、CAPture、SMART、CAP-trapper、CapSelect、CAP-jumping。这几种方法在起始材料用量、文库构建技术、实验操作复杂程度和文库构建质量等方面构存在不同程度的优缺点,但综合比较而言,SMART和CAP-trapper这两种方法比较有实际应用价值。SMART法适于简单、快速地构建一般质量的cDNA文库,如果采用Little-cycleSMART和Large-sizeSMART这两种改进技术,所建文库质量也非常好;CAP-trapper法虽然对实验技术要求较高,实验过程复杂,但所建文库全长率高,冗余性低,建库效率高,是目前构建高质量文库最好的方法。

The full length cDNA library is regarded as an economical, rapid and effective way to functional genomic research, it welcomes the shortcoming and save the time of gene cloning and function identifying result in accelerating the research of functional genomics. At present, the methods of constructing the full length cDNA library include Oligo-Capping, CAPture, SMART, CAP-trapper, CapSelect and CAP- jumping method.. To some extend, these methods exist their own advantages and disadvantages in the amount of start materials ,the technology and process of experiment and the quality of the cDNA library. It is compared that the value of the SMART and CAP-trapper method are relatively higher in utility. The SMART method is adapt to construct easily and rapidly cDNA library with average quality. The quality of the library can be improved, if the Little-cycle SMART and Large-size SMART technology are used to construct the full length cDNA library. The CAP-trapper method is adapt to construct cDNA library with high complement rate,low repeat rate and high effiency, as the technology is high and the process is complex.