Sample remove place isolation add measure required practical

Required Practical 8 investigates dehydrogenase activity in chloroplasts by grinding leaves, isolating chloroplasts via centrifugation, and measuring the absorbance of DCPIP over time at different light intensities using a colorimeter.

Vereist Praktisch 8: Onderzoek naar het effect van een benoemde factor op de activiteitssnelheid van dehydrogenase in extracten van chloroplasten 1. Verwijder stengels van bladmonsters. Maal monster met vijzel en stamper en plaats in een gekoelde isolatieoplossing. 2. Gebruik een kaasdoek en trechter om het monster in een beker te filteren. Plaats de beker in een ijs-waterbad om het monster koel te houden. 3. Breng over naar centrifugebuizen en centrifugeer 10 minuten op hoge snelheid. Dit scheidt chloroplasten in de pellet. 4. Verwijder het supernatant en voeg het pellet toe aan vers isolatiemedium. Bewaar isolatieoplossing op ijs. 5. Stel de colorimeter in op het rode filter. Nulstellen met een cuvette met chloroplastenextract en gedestilleerd water. 6. Plaats de reageerbuis 30 cm van de lichtbron en voeg DCPIP toe. Neem onmiddellijk een monster en doe dit in de cuvette. Meet de absorptie van het monster met de colorimeter. 7. Neem elke 2 minuten gedurende 10 minuten een monster en meet de absorptie. 8. Herhaal voor verschillende afstanden van de lamp tot 100 cm. Dit varieert de lichtintensiteit.

Text	Required Practical 8: Investigation into the effect of a named factor on the rate of dehydrogenase activity in extracts of chloroplasts 1. Remove stalks from leaf samples. Grind sample using a pestle and mortar and place into a chilled isolation solution. 2. Use a muslin cloth and funnel to filter the sample into a beaker. Suspend the beaker in an ice water bath to keep sample chilled. 3. Transfer to centrifuge tubes and centrifuge at high speed for 10 minutes. This will separate chloroplasts into the pellet. 4. Remove supernatant and add pellet to the fresh {{c4:isolation medium}}. Store isolation solution on ice. 5. Set the colorimeter to the red filter. Zero using a cuvette containing chloroplast extract and distilled water. 6. Place test tube in the rack 30cm from light source and add DCPIP. Immediately take a sample and add to cuvette. Measure the absorbance of the sample using the colorimeter. 7. Take a sample and measure its absorbance every 2 minutes for 10 minutes. 8. Repeat for different distances from lamp up to 100 cm. This will vary the light intensity.

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Required Practical 8: Investigation into the effect of a named factor on the rate of dehydrogenase activity in extracts of chloroplasts

1. Remove stalks from leaf samples. Grind sample using a pestle and mortar and place into a chilled isolation solution.

2. Use a muslin cloth and funnel to filter the sample into a beaker. Suspend the beaker in an ice water bath to keep sample chilled.

3. Transfer to centrifuge tubes and centrifuge at high speed for 10 minutes. This will separate chloroplasts into the pellet.

4. Remove supernatant and add pellet to the fresh {{c4:isolation medium}}. Store isolation solution on ice.

5. Set the colorimeter to the red filter. Zero using a cuvette containing chloroplast extract and distilled water.

6. Place test tube in the rack 30cm from light source and add DCPIP. Immediately take a sample and add to cuvette. Measure the absorbance of the sample using the colorimeter.

7. Take a sample and measure its absorbance every 2 minutes for 10 minutes.

8. Repeat for different distances from lamp up to 100 cm. This will vary the light intensity.

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