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Buffers and Solutions

Agarose solutions.
Ethidium bromide.
Electrophoresis buffer.

Nucleic Acids:
DNA samples.
DNA Ladders.

Preparation for 50 mL
 Prepare a 50x stock solution of TAE buffer
 Prepare sufficient electrophoresis buffer (1x TAE)
 Prepare a solution of agarose in electrophoresis buffer at an appropriate concentration

1. Prepare TAE buffer.

2. Weigh 1 grams of agarose and add to the 50 mL of disilled water + 1 mL 50x TAE buffer.

3. Keep in oven.

4. Take the solution from oven.

5. Add 1 μl of ethidium bromide.

6. Pour the solution to a gel caster.

7. Place the comb.

8. Pour the 100ml 1x TAE buffer solution to the electrophoretic chamber.

9. Place the gel in the caster in the electrophoretic chamber.

10. Connect the electrodes and switch on the current.

11. Switch off the power supply.

12. Remove the gel from the electrophoretic chamber.

13. Place the gel in the UV Transilluminator.

14. Switch on the Transilluminator.

0 ml of 75% ethanol (invert tube) Centrifugation for 4.000 g for 2 mins DNA wash (2x) Wash the DNA precipitate 0. Lysis/Homogenization 1 ml DNAzol® Reagent + cell samples 2. DNA Solubilization 8 mM NaOH ethanol and 8 mM NaOH Get cell pellet of bacteria culture by centrifugation at 3.000 g for 2 mins Remove supernatant DNA Solubilization Air dry DNA by an open tube < 1 min after removing the ethanol Dissolve the DNA in 50-100 μl 8 mM NaOH Addition of 15.8-1.000 × g.DNA extraction by DNAzol Protocol 1. 10 min 3. DNA Precipitation Lysate + 0. DNA Wash 1 ml 75% ethanol 5.000 rpm for 5 mins Lysis/Homogenization Mix Cell pellet + 1 mL of DNAzol by gently pipetting Centrifugation Centrifugation at 10. Centrifugation (optional) 10.000 g for 10 mins Get supernatant (transfer to fresh tube) DNA Precipitation Supernatant + 0.5 ml 100% ethanol 4.9 μl 0.1 M HEPES .5 mL of 100% ethanol (invert tube) Store at room temperature for 3 mins Centrifugation for 4.