Instructions

DNA Extraction from Kiwi fruit - please watch first

Step 1

Extract Kiwi pulp (detergent, salt and water mix) as shown in the video

Heat extract 15' at 60°C

Cool and filter

Take a sample (0.5ml) of the filtrate and place it in an Eppendorf tube

Add 1ml cold EtOH and leave on ice for 15 minutes

Place the tube in a microcentrifuge (ensure that there is an appropriate balance tube) and spin on maximum speed for 15 minutes

Remove the supernatant as much as you can.

Add 1 mL of water to the pellet, suspend, spin again

Transfer the supernatant into a suitable cuvette

Measure the supernatant in the virtual Spectrophotometers at 260 and 280 nm below

Record your measurements

Prepare 3 Eppendorf tubes and label them F1, F2 and F3.

Add 50 µl fish DNA (1 mg/mL) to each of the tubes.

To tube F1 add 100 µl 1M NaCl and 0.35 ml water, mix well and then add 1ml cold EtOH

To tube F2 omit the salt add 0.45 ml water, mix well, then add add 1 ml cold EtOH)

To tube F3 add 0.95 ml water, mix well and set aside for later

Put all three tubes on ice and leave them for 15 minutes

Spin F1 and F2 (not F3) and resuspend pellets in 1ml water

Place tubes F1 and F2 (not F3) in a microcentrifuge (ensure that there is an appropriate balance tube) and spin on maximum speed for 15 minutes

Remove the supernatant as much as you can.

Add 1 mL of water to the pellet, suspend, spin again

Transfer the supernatant into a suitable cuvette and measure the absorbance for F1 - F3 at 260 nm and 280 nm.

Measurements for the samples with fish DNA:

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