CONSTRUCTION AND CHARACTERIZATION OF GOLDEN HAMSTER NEURAL TUBE cDNA LIBRARY / 解剖学报

ABSTRACT

Objective Constructing golden hamster neural tube cDNA library in order to detect new genes associated with development of neural tube and neural tube defects. Methods Total RNAs were isolated from neural tube using TRIZOL Reagent; Doublestranded cDNAs were synthesized from total RNAs using LD-PCR method; The dsc DNAs were digested by proteinase K and Sfi Ⅰ, less than 500?bp fragments were separated by CHROMA SPIN-400, and then the cDNAs were ligated to ? TriplEX-2 vector.After packaging,the neural tube cDNA library was constructed. Results The titer of neural tube cDNA library was 1.5?10 6?pfu/ml.The recombination rat was 99%.The average insert was larger than 1?kb.Conclusion Our successfully constructed cDNA library is a full-length library with high efficiency,and can be used to detect the genes related to development of neural tube and neural tube defects.