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Construction of low melting point agarose emulsion PCR amplification complex gene sequence
Author(s) -
Minxuan Xu,
Bingchang Tan,
Wei Zhou,
Ting Wei,
Honglin Zhang,
Dongrui Zhou
Publication year - 2012
Publication title -
journal of genetic engineering and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.729
H-Index - 25
eISSN - 2090-5920
pISSN - 1687-157X
DOI - 10.1016/j.jgeb.2011.12.001
Subject(s) - agarose , emulsion , template , dna , polymerase chain reaction , chemistry , plasmid , microbiology and biotechnology , chromatography , gene , biology , materials science , nanotechnology , biochemistry
Emulsion polymerase chain reaction, an effective amplification, can make millions of templates could be individually amplified within a single tube. Here we constructed and improved a low melting point agarose-emulsion method to promote the specific sequences amplification effectively. Artificial Lactobacillus Plasmid as template was amplified and clear fluorescence images of the agarose beads were obtained. The Real-time PCR data showed that agarose-emulsion PCR clearly indicated that DNA can be amplified in agarose droplets. Overall, our study effectively overcame the difficulty of formation of uniform emulsion droplets, negative effect on recombination of homologous regions of DNA and generation of void emulsion droplets. This method increases the accuracy with amplification, reduces the influence of uncertainties and provides the reliable data for further experiment

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