Isolation and restriction analysis of chloroplast DNA from Hevea


Citation

Cheong K.F., . and Chew N.P., . and Koh C.L., . 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 DNA (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 electrophoretogram 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 XhoI-generated fragments wa 152.3 kbp


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Abstract

A procedure for the isolation and purification of chloroplast DNA (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 electrophoretogram 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 XhoI-generated fragments wa 152.3 kbp

Additional Metadata

[error in script]
Item Type: Article
Additional Information: Summary (En)
AGROVOC Term: HEVEA BRASILIENSIS
AGROVOC Term: PURIFICACION
AGROVOC Term: CLOROPLASTO
AGROVOC Term: ADN
AGROVOC Term: GENOMAS
AGROVOC Term: PLANTAS LACTIFERAS
Depositing User: Ms. Norfaezah Khomsan
Last Modified: 24 Apr 2025 05:56
URI: http://webagris.upm.edu.my/id/eprint/20911

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