Team:Evry/Protocols/15
From 2013.igem.org
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Put the solution into microwave until Agarose is disolved.</br> | Put the solution into microwave until Agarose is disolved.</br> | ||
When until the cool down and add 3 mL of Ethidium Bromide (EtBr).</br> | When until the cool down and add 3 mL of Ethidium Bromide (EtBr).</br> | ||
- | Put the solution in the cuve and let it cool down. < | + | <FONT COLOR=#ff0000 ><b>EtBR is a carcinogen substance, use it with caution !</FONT></b><br/> |
+ | Put the solution in the cuve and let it cool down. <br/></p> | ||
<p><b>Mix gel</p></b> | <p><b>Mix gel</p></b> | ||
Put 5 μL of PCR product and 1 μL of loading dye 6X in each well.<p> | Put 5 μL of PCR product and 1 μL of loading dye 6X in each well.<p> | ||
- | Do not forget your positive and negative controles, and the kB ladder.< | + | Do not forget your positive and negative controles, and the kB ladder.<br/> |
Put in the electrophoresis cell at 100 V during 40 minutes. </p> | Put in the electrophoresis cell at 100 V during 40 minutes. </p> | ||
Latest revision as of 11:58, 6 September 2013
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