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Published 20 October 2013
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DNA-Fingerprint - Procedure

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Procedure of DNA-Fingerprints

Tubes of each work group:

1. Preparation of the Restriction
1.1. Pipet 10 µl of enzyme mix into each tube.

2. Restriction Digestion
Place the tubes in the floating rack and incubate for 20 min at 37 °C in a water bath.

3. Prepare Gel Electrophoresis
3.1. Preparing the electrophoresis chamber
3.2. Produce the gels:
- Give 0,24 g agarose + 30 ml TAE-buffer into a 250 ml Erlenmeyer flask.
- Boil the Erlenmeyer flask shortly in the microwave, swirl it and boil it shortly again.
3.3 Filling the chamber
- Pour the gel at 55 °C into the gel box. Test the temperature on the back of your hand.
- After the gel has become solid, remove the metal rods of the electrophoresis chamber.
- Fill the chamber with 270 ml of TAE-buffer to cover the gel.
- Pull the comb.

4. Preparation of DNA marker
Pipet 10 µl loading buffer (LB) into 10 µl DNA marker (M).

5. Addition of 10 µl loading buffer
Pipet 10 µl of the loading buffer into the CS and S1 – S5.

6. Filling the wells
Pipet 20 µl of each sample and 20 µl DNA marker (M) into the wells.
Remember the sequence!

7. Gel elektrophoresis (120 V)
Stop the electrophoresis when the blue band has migrated to 1 cm before the end of the gel.

8. Making DNA visible
- Transfer the gel into the colouring bowl and cover it with 60 ml of colouring solution.
- Put the colouring solution back into the Erlenmeyer flask.

9. Analyse the gel
Which of the suspects is the perpetrator?

There are two files of the procedure of DNA-fingerprint practice course, in pdf and word document and a ppt presentation of this experiment.