Team:Evry/Protocols/15

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<p>Electrophoresis is a method using electrical field to separate DNA or RNA sequence by size. Smaller the fragment is, the more it migrates on the gel. <br>
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<p>After <a href='https://2013.igem.org/Team:Evry/Protocols/07' target='_blank'>PCR</a>, 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. <br>
Using a DNA ladder, we can know the size of DNA sequence and then check if we have the sequence we wanted.
Using a DNA ladder, we can know the size of DNA sequence and then check if we have the sequence we wanted.
</p>
</p>
<h2> Preparation</h2>
<h2> Preparation</h2>
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<p><b>Agarose gel 1% preparation:</b><br/>
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Add 0,5 g of Agarose in 50 mL of TAE 1X </br>
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(See protocole for TAE 1X preparation <a href='https://2013.igem.org/Team:Evry/Protocols/04' target='_blank'>there</a>)</br>
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Put the solution into microwave until Agarose is disolved.</br>
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When until the cool down and add 3 mL of Ethidium Bromide (EtBr).</br>
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<FONT COLOR=#ff0000 ><b>EtBR is a carcinogen substance, use it with caution !</FONT></b><br/>
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Put the solution in the cuve and let it cool down. <br/></p>
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<p><b>Mix gel</p></b>
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Put 5 μL of PCR product and 1 μL of loading dye 6X in each well.<p>
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Do not forget your positive and negative controles, and the kB ladder.<br/>
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Put in the electrophoresis cell at 100 V during 40 minutes. </p>
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<h2> Analysis</h2>
<h2> Analysis</h2>

Latest revision as of 11:58, 6 September 2013

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

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