Citation
Cheong K. F., . and Koh C. L., . and Chew N. P., . Isolation and restriction analysis of chloroplast DNA from Hevea. pp. 278-288. ISSN 0127-7065
Abstract
A procedure for the isolation and purification of chloroplast DMA (ctDNA) from Hevea is described. Differential centrifugation of Hevea leaf homogenates separates most of the plastids from nuclei. By means of sucrose density gradient centrifugation chloroplasts of high integrity have been obtained. These chloroplasts were apparently nuclei-free when observed by phase-contrast microscopy. DNA was prepared and purified from the isolated chloroplasts by organelle lysis and density gradient isopycnic centrifugation. The purified ctDNAs obtained were amenable to digestions with restriction endonucleases. The low background of UV-fluorescent material in the electrophoretograms of restricted Hevea ctDNA was consistent with minimal nuclear DNA contamination in the ctDNA preparations. The average genome size of Hevea ctDNA as estimated by summation of HindIII- PvuII- SalI- or Xhol-generated fragments was 152.3 kbp.
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Abstract
A procedure for the isolation and purification of chloroplast DMA (ctDNA) from Hevea is described. Differential centrifugation of Hevea leaf homogenates separates most of the plastids from nuclei. By means of sucrose density gradient centrifugation chloroplasts of high integrity have been obtained. These chloroplasts were apparently nuclei-free when observed by phase-contrast microscopy. DNA was prepared and purified from the isolated chloroplasts by organelle lysis and density gradient isopycnic centrifugation. The purified ctDNAs obtained were amenable to digestions with restriction endonucleases. The low background of UV-fluorescent material in the electrophoretograms of restricted Hevea ctDNA was consistent with minimal nuclear DNA contamination in the ctDNA preparations. The average genome size of Hevea ctDNA as estimated by summation of HindIII- PvuII- SalI- or Xhol-generated fragments was 152.3 kbp.
Additional Metadata
| Item Type: | Article |
|---|---|
| AGROVOC Term: | Hevea brasiliensis |
| AGROVOC Term: | Chloroplasts |
| AGROVOC Term: | Isolation |
| AGROVOC Term: | DNA |
| AGROVOC Term: | Organelles |
| AGROVOC Term: | Centrifugation |
| AGROVOC Term: | Plastids |
| AGROVOC Term: | Microscopy |
| AGROVOC Term: | Purification |
| AGROVOC Term: | Genomes |
| Geographical Term: | Malaysia |
| Depositing User: | Ms. Suzila Mohamad Kasim |
| Last Modified: | 28 Apr 2025 04:31 |
| URI: | http://webagris.upm.edu.my/id/eprint/23425 |
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