Describe how a sample of whole chloroplasts could be obtained from leaf tissue
please could someone point me in the right direction
what group is a frog in
Since this is not my area of expertise, I searched Google under the key words "leaf tissue chloroplasts sample" to get this possible source:
http://www.sigmaaldrich.com/img/assets/13700/organelle_isolation.pdf
In the future, you can find the information you desire more quickly, if you use appropriate key words to do your own search.
I hope this helps. Thanks for asking.
To obtain a sample of whole chloroplasts from leaf tissue, you can follow the steps below:
1. Obtain fresh leaf tissue: Start by selecting healthy leaves from the plant of interest. Make sure the leaves are young and green, as mature leaves tend to have fewer chloroplasts.
2. Cut leaf tissue into small pieces: Use a sharp blade or scissors to cut the leaf into small pieces. This will increase the surface area and make it easier to extract the chloroplasts.
3. Isolate leaf cells: Gently grind the leaf tissue in a mortar and pestle with a small amount of extraction buffer solution. The extraction buffer typically contains chemicals that help in breaking down the cell wall and releasing the chloroplasts. The buffer can vary based on the research protocol or intended use of the chloroplasts.
4. Filter the mixture: After grinding, transfer the leaf tissue extract to a test tube or centrifuge tube. To remove large debris and cell fragments, filter the extract using a fine mesh or cheesecloth.
5. Centrifuge the extract: Spin the filtered extract in a centrifuge at a relatively low speed to separate chloroplasts from other cellular components. With centrifugation, the heavier chloroplasts will pellet at the bottom of the tube while other debris remains in the supernatant.
6. Resuspend and wash the chloroplasts: Carefully remove the supernatant and gently resuspend the pellet of chloroplasts in a fresh buffer solution. This will help to wash away any remaining contaminants or debris.
7. Repeat centrifugation and resuspension: Repeat the centrifugation step to further purify the chloroplasts. The more times you repeat this process, the purer your chloroplast sample will become.
8. Collect and store the chloroplasts: After the final centrifugation and resuspension, carefully remove the supernatant and collect the purified chloroplasts from the pellet. Transfer them to a fresh tube and store in a suitable storage solution.
It is important to note that the specific protocol and buffer components used may vary depending on the purpose of the chloroplast isolation and the specific research methods being employed. Always refer to established protocols or consult with experts for the most accurate and reliable results.