Team:Evry/Protocols/15

From 2013.igem.org

Iron coli project

Gel electrophoresis analysis

Principle

After PCR, electrophoresis is a method using electrical field to separate DNA or RNA sequences by size. Smaller the fragment is, the more it migrates on the gel.
Using a DNA ladder, we can know the size of DNA sequence and then check if we have the sequence we wanted.

Preparation

Agarose gel 1% preparation:
Add 0,5 g of Agarose in 50 mL of TAE 1X
(See protocole for TAE 1X preparation there)
Put the solution into microwave until Agarose is disolved.
When until the cool down and add 3 mL of Ethidium Bromide (EtBr).
EtBR is a carcinogen substance, use it with caution !
Put the solution in the cuve and let it cool down.

Mix gel

Put 5 μL of PCR product and 1 μL of loading dye 6X in each well.

Do not forget your positive and negative controles, and the kB ladder.
Put in the electrophoresis cell at 100 V during 40 minutes.

Analysis