Tissue selection and the optimization of the DNA isolation method for the construction of genomic library of red dragon fruit (Hylocereus polyrhizus) / Daniel Lee Kian Poh
Genomic library of Hylocereus polyrhizus was constructed in bacteriophage Lambda Fix II with 20kb inserts. DNA of the H. polyrhizus that was used to construct the genomic library was obtained from young germinated seeds’ leaves. The tender and partially expended leaves were selected. Seed germina...
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Format: | Thesis |
Published: |
2013
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Online Access: | http://studentsrepo.um.edu.my/4141/1/SGR080162_Daniel_Lee_Kian_Poh.pdf http://pendeta.um.edu.my/client/default/search/detailnonmodal/ent:$002f$002fSD_ILS$002f988$002fSD_ILS:988338/ada?qu=Tissue+selection+and+the+optimization http://studentsrepo.um.edu.my/4141/ |
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Summary: | Genomic library of Hylocereus polyrhizus was constructed in bacteriophage Lambda
Fix II with 20kb inserts. DNA of the H. polyrhizus that was used to construct the
genomic library was obtained from young germinated seeds’ leaves. The tender and
partially expended leaves were selected. Seed germination was also carried out to select
the best time to harvest leaves for DNA extraction. Different treatment for seeds affects
the germination and growth of the seed and acid washed seeds germinated in light
condition the best. DNA extraction was modified in order to obtain pure and high yield
DNA for genomic library construction. Restriction enzyme (BamH I) used to digest the
selected fragment from the total DNA and analysed on a gel electrophoresis. Phenolchloroform
method used to purify insert DNA successfully yielded high concentration
insert DNA which is a very crucial step in constructing genomic library. Plaques formed
indicated that the bacteriophage has successfully lysised the cells. In this study, 80 –
100 plaques per Petri dish formed showed bacteriophage has successfully lysised the
cells and indicated that a good genomic library has been established. This genomic
library can be use for further study such as fish out important gene and study the gene
interaction. |
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