Screen for recombinant clones by colony Polymerase chain reaction
Xiu-jie Sheng
Abstract
Xiu-jie Sheng
Abstract
Objective To screen the Doc 1R gene recombinant plasmid by use of colony PCR. Method The recombinant colonies were transfered into the PCR reaction mixture. The PCR primers were used for constructing mouse Doc 1R genomic sequence. Result Among the 5, 3 positive strips in the size of 1 500 bp were visible, which were the same as the Doc 1R gene fractions in terms of their sizes were screened as positive clones. The positive colony were further confirmed by double digestion and DNA sequencing. Conclusion Colony PCR is a simple, efficient and reliable technique for screening the recombinant.
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Objective To screen the Doc 1R gene recombinant plasmid by use of colony PCR. Method The recombinant colonies were transfered into the PCR reaction mixture. The PCR primers were used for constructing mouse Doc 1R genomic sequence. Result Among the 5, 3 positive strips in the size of 1 500 bp were visible, which were the same as the Doc 1R gene fractions in terms of their sizes were screened as positive clones. The positive colony were further confirmed by double digestion and DNA sequencing. Conclusion Colony PCR is a simple, efficient and reliable technique for screening the recombinant.
Key concepts: Recombinant DNA, Polymerase chain reaction, Molecular biology, genomic DNA, Biology, Plasmid, Gene, DNA sequencing