Studies On PicoPlanktonic Eu-Bacteria 16s RDNA Character Of RFLP And Relevant Methods In Jiaozhou Bay

The diversity of picoplanktonic eubacteria were investigated by comparing the RFLP (restriction fragment length polymorphism) of 1 6s rDNA through the methods introduced from molecular biology. There are at least seven kinds of eubacteria in Jiaozhou Bay differed in the digested 1 6s rDNA fragment by EcoR I and Hind III. Also some relevant methods are constructed. First, successful extraction and purification of bacterial genomic DNA from sea water proved excellent. There is O.3----O.5 ~?g DNA available from per litre sea water. Seconds Competent cells (DH5 a ) are obtained by the process of CaC12 and SSCS solution. It is obvious SSCS solution simpler process and higher efficiency than CaC12. Third, using the two proper primers, Universal 1406R and Eubacterial 68F,about 90% 16s rDNA in bacteria was enlarged by appropriate PCR(polymerase chain reaction) condiction. Forth, PCR production was successfully ligated with pUCm桾 plasmid by TA cloning and transformed into competent E.Coli cells This thesis would be helpful to construct eubacterial phylogenetic analysis tree in Jiaozhou bay ,and the methods are easily used in other materials, such as Archaea, Prochlorococcus.