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Whole-ceilPCR to identify Anabaena species and strains
Huang Jia-Quan; Wang Gao-Hong; Li Dun-Hai; Liu Yong-ding (liuyd@ihb.ac.cn); Yin Li-Yan; Inst Hydrobiol, Chinese Acad Sci, Wuhan, 430072, China
2004
Source PublicationActa Hydrobiologica Sinica
ISSN1000-3207
Volume28Issue:2Pages:159-162
AbstractThe traditional way to identify algae is usually depend on the observation of the morphology. This way is time consuming and usually difficult to identify the algae during the early algal growth stage in which the character is not obvious and can not detect the subtle differences between species or strains; also, the morphology and the ability to produce toxicants of cyanobacteria may change under different natural or cultural conditions. In this paper, we use the molecular method to identify Anabaena species, the results showed that BSA are necessary for the PCR amplification and the optimal concentration is 1% (w/v) in the reacion buffer. DMSO increased or decreased the quantity of the amplification products selectively, but the effect is not significant. The self designed primers after comparison of other Anabaena PC sequences amplified the part of pcb and pca gene along with PC-IGS sequence when using both extracted DNA and pretreated whole cells as template, the banding pattern is not uniform but all produced the target bands, cloning and sequencing analyses indicated that PC in line with PC-IGS was amplified, even though the nucleotide between two Anabaena strains collected from two different regions in China show that only 2 sites in a fragment about 400 bp are different, but the PC-IGS sequences are quite different from others and thus can be applied to identify the different species or strains. The universal primers to amplify part of 16S rDNA and 23S rDNA along with their ITS sequence using the extracted DNA and pretreated whole cell showed that the banding pattern of amplification products are uniform when the whole cells were at the concentration of 105/mL in the reaction buffer, the minimum amount of whole cell to amplify the target band is 20 cells in 50muL reaction buffer.
Indexed By其他
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/18133
Collection期刊论文
Corresponding AuthorHuang Jia-Quan; Inst Hydrobiol, Chinese Acad Sci, Wuhan, 430072, China
Recommended Citation
GB/T 7714
Huang Jia-Quan,Wang Gao-Hong,Li Dun-Hai,et al. Whole-ceilPCR to identify Anabaena species and strains[J]. Acta Hydrobiologica Sinica,2004,28(2):159-162.
APA Huang Jia-Quan,Wang Gao-Hong,Li Dun-Hai,Liu Yong-ding ,Yin Li-Yan,&Inst Hydrobiol, Chinese Acad Sci, Wuhan, 430072, China.(2004).Whole-ceilPCR to identify Anabaena species and strains.Acta Hydrobiologica Sinica,28(2),159-162.
MLA Huang Jia-Quan,et al."Whole-ceilPCR to identify Anabaena species and strains".Acta Hydrobiologica Sinica 28.2(2004):159-162.
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