Chloroplasts Essay

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Chloroplast fractionation: Nucleic acid and protein analysis via gel electrophoresis
ABSTRACT: Chloroplasts carry out photosynthetic processes to meet the metabolic demands of plant cells (Alberts, 2008). They consist of an inner thylakoid membrane and a stroma. (Parent et. al, 2008).In this experiment we demonstrate the unique protein compositions of isolated thylakoid and stromal fractions from broken and whole spinach chloroplasts. Because these compartments carry out different metabolic processes, we confirm our hypothesis that performing SDS-PAGE on these fractions will result in distinct patterns on the gels. In isolating and analyzing nucleic acid from broken, whole, and crude chloroplast samples we demonstrate that genes for photosynthetic protein psbA are found in chloroplast DNA, while genes for photosynthetic enzyme …show more content…

These six samples (crude -/+, broken -/+, and whole -/+) were spun at 5000 rpm, and the resulting pellets were isolated and resuspended in DNase buffer. The set of suspensions labeled with a (+) was incubated in DNase enzyme for 15 minutes, and afterwards incubated in 15 uL of STOP solution. All six samples were lysed for DNA extraction with DNA extraction buffer, and micro-centrifuged at maximum speed. To precipitate the extracted DNA, the supernatants from each of the six samples were added to their correspondingly labeled micro-centrifuge tubes containing 7% ethanol (Parent et. al, 2008To bind the DNA, the ethanol lysate mixtures were transferred to labeled spin columns and spun for one minute in the micro-centrifuge at maximum speed. To wash the bound DNA, the spin columns were washed and spun three times at maximum speed. In order to elute the bound DNA, the samples were washed in 80 uL of distilled water and spun again for 2 minutes at maximum speed (Parent et. al,

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