The intent of the survey is to gauge the familial fluctuation and species distinction harmonizing to geographical differences of Hygrophila pogonocalyx by utilizing RAPD ( random amplified polymorphous DNA ) fingerprinting, which is a PCR-based fingerprinting method. With the survey of the workss familial fluctuations and phylogeography, we can acquire a better position on the workss evolutionary history and can happen solutions to protect the endangered species.
H. pogonocalyx is an aquatic works which is badly threatened due to human activities. In Taiwan, there are four populations staying separated by geographical differences which is the mountain ridge. As a consequence, familial fluctuation between the populations is important. From the above observations, population distinction is influenced by phylogeography, and therefore there are familial impetuss among the works populations. Besides from geographical differences, pollinators such as insects will besides act upon the familial variableness of a population. This is due to the fact that some insects will merely be localized at a specific country. In add-on, some seeds dispersion of workss will besides limited to a certain country.
From the above observation, RAPD analysis can be used to analyze the differences and similarities between the populations of H. pogonocalyx. RAPD is a polymerase concatenation reaction ( PCR ) -based method which was used to magnify H. pogonocalyx DNA. By utilizing the RAPD fingerprinting, we can find that there is familial diverseness nowadays as it functions as a familial marker to observe familial fluctuations among strains of H. pogonocalyx.
MATERIALS AND METHODS
H. pogonocalyx was collected from northeasterly and western Taiwan. As there are merely limited populations available, merely 10 % is collected from each population and this will do certain that the same person is non collected twice. The samples collected must be placed in a cool country and is cultured. Before continuing to the grinding procedure, the foliages of immature shoots are washed and the grinded foliage tissue pulverization was placed in liquid N to interrupt down cell wall stuff and let entree to DNA while harmful cellular enzymes and chemicals remain inactivated. The genomic Deoxyribonucleic acid of the works was extracted by utilizing the works DNA extraction method. The Deoxyribonucleic acid was resuspended in CTAB buffer. To sublimate the Deoxyribonucleic acid, centrifugation was used to take indissoluble particulates and other stuffs were separated by blending with trichloromethane and centrifuged. The supernatant was transferred and was centrifuged once more and the pellet was washed by ethyl alcohol to take polluting salts. The pellet was so resuspended in TE buffer. The genomic Deoxyribonucleic acid extracted was run on agarose gel, stained with ethidium bromide and observed under UV visible radiation.