Unit cloning and amplification as novel and universal strategies for complex vector construction and small DNA fragment preparation.
With a novel and universal strategy for the cloning of multiple DNA fragments, a complex synthetic vector (pVEC100), harboring the target DNA fragments in conventional 100-bp DNA ladder, was constructed for efficient and large-scale production of 100-bp DNA marker through bacteria fermentation, plasmid extraction and restrictive digestion. Since the restrictive digestion of complex vectors yields insufficient small DNA fragments, an innovative PCR model was developed as an alternative. The PCR model comprised a specially designed template vector and a unit amplification model for producing groups of small DNA fragments. The unit amplification model improved the efficiency of the PCR protocol and made it more economical and easier for small DNA fragment amplification. The approach presented in this paper--a unit cloning model for constructing complex synthetic vectors combined with the modular design of unit amplification by PCR--is a powerful method for preparing small DNA fragments of DNA molecular weight standards.
Ye
Chunjiang
C
Biotechnology Department, The School of Medical and Life Science, University of Jinan, Jinan, P. R. China. chm_yecj@ujn.edu.cn
Gu
Jingsong
J
Chen
Sunxiao
S
Deng
Anmei
A
Li
Yue Zhong
YZ
Li
Dianxiang
D
D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D004587:Electrophoresis, Agar Gel / Q000592:standards*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018745:Genome, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards
D004247:DNA; D015246:Deoxyribonuclease EcoRI