Team:Grenoble/Biology/Protocols/Gel

From 2012.igem.org

(Difference between revisions)
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         <li>Add 1 g of agarose powder to 50 mL of 1X TAE buffer.</li>
         <li>Add 1 g of agarose powder to 50 mL of 1X TAE buffer.</li>
         <br/>
         <br/>
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         <li>Heat the solution using the microwave until the powder is completely dissolved.</li>
+
         <li>Heat the solution using the microwave until the agarose is completely dissolved.</li>
         <br/>
         <br/>
         <center>
         <center>
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                 <tr><th><i><div class="petit">SAFETY AND USEFUL RECOMMANDATION</div></i></th></tr>
                 <tr><th><i><div class="petit">SAFETY AND USEFUL RECOMMANDATION</div></i></th></tr>
                 <tr><td><div class="petit">When heating the mixture it is important to use an adaptable container.
                 <tr><td><div class="petit">When heating the mixture it is important to use an adaptable container.
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                 Putting metallic things in the microwave is dangerous. It causes the formation of intense electric
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                 Putting metallic objects in the microwave is dangerous. It causes the formation of intense electric
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                 current. Actually falcon tubes are used. But there must be left open, because the elevation of
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                 current. Keep in mind that the container might explode causing the deterioration of the microwave, the preparation and possible injuries to the  
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                temperature can make the pressure increase. And if the tubes are not open, these one could explose,
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people next to the device.  
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                causing the deterioration of the microwave and of the preparation. It also can burn people. What is
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To avoid this kind of situation the microwave is only used for short heating periods with the user keepin a eye on the preparation. </div></td></tr>
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                done to prevent this situation is that the microwave is turned off every 2 minutes to see if the
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                preparation is good or not. </div></td></tr>
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             </table>
             </table>
         </center>
         </center>
         <br/>
         <br/>
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         <li>Once the solution is cool enough to be touched, pour it into gel cast (don't forget the comb).</li>
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         <li>Once the solution's temperature has decreased (enough to be able to touch the container), pour it into a gel cast (don't forget the comb).</li>
</section>
</section>
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     <h2>Load the gel</h2>
     <h2>Load the gel</h2>
     <ol>
     <ol>
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         <li>When the gel is solid, remove carefully the comb.</li>
+
         <li>When the gel is solid, carefully remove the comb.</li>
         <br/>
         <br/>
         <li>Place the gel in the buffer tank and cover it with the 1X TAE buffer solution.</li>
         <li>Place the gel in the buffer tank and cover it with the 1X TAE buffer solution.</li>
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         <li>Chose the right voltage.</li>
         <li>Chose the right voltage.</li>
         <br/>
         <br/>
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         <li>Put the device on.</li>
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         <li>Turn on the device.</li>
         <br/>
         <br/>
         <center>
         <center>
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                 <tr><td><div class="petit">The generator can deliver over 130V. The risk of electrocution
                 <tr><td><div class="petit">The generator can deliver over 130V. The risk of electrocution
                 for people is generally around 50V. That is why some recommendations must be followed.
                 for people is generally around 50V. That is why some recommendations must be followed.
-
                 First before using the electrophoresis device, users should check if they have all parts
+
                 Before using the electrophoresis device, users should check if they have all parts
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                 of the device, because it is a removable one. They also have to check that each part
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                 of the device, because they are removable. They also have to check that each part
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                 corresponds to the good device. Indeed there are different kinds of tank and using parts
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                 corresponds to the right device. There are different kinds of tanks and parts are not compatible with each other.
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                 of different devices will not make the device function safely. That is why we managed to
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                 Using parts from another type of tank may lead to the loss of safety protections. We managed to assign a
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                 number each part in order to know from which device they belong to. Then the generator must
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                 number to each corresponding part in order to know to which device they belong to. The generator must
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                 not be in presence of water and its vents must not be blocked, because the device will become
+
                 not be in presence of water and its vents must not be blocked. The generator has two main safety protections: a detector of migration buffer and a lid.  
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                hot during its use. Moreover the generator has two main safety protections. The first one
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                The detection does not let the device generate electricity until the migration buffer has been put in the tank and reached the correct level.  
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                is a detection of migration buffer. This detection does not let the device generate electricity
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                This is a protection both for the device and for users. Moreover, it prevents
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                until the migration buffer has been put in the tank and has reached a correct level. This is a
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                 users to be surprised by an unexpected production of electricity during the manipulation. The second protection prevents the
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                protection both for the device and for users. Thanks to this protection the device could not
+
-
                generate electricity when something was not be done correctly by users. Moreover it also prevents
+
-
                 users to be surprised by the production of electricity during the manipulation. The second
+
-
                protection is a mechanic one. Thus it seems to be more efficient. This protection prevents the
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                 generator to produce electricity when no lid is put on the tank. With this, there is less risk of
                 generator to produce electricity when no lid is put on the tank. With this, there is less risk of
-
                 injuries and accidents. But it should be use correctly after all.</div></td></tr>
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                 injuries and accidents, if it's correctly used.</div></td></tr>
             </table>
             </table>
         </center>
         </center>
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Revision as of 10:15, 29 August 2012

iGEM Grenoble 2012

iGEM Grenoble iGEM
iGEM 2012
main page
Project

Gel electrophoresis

Goal

Separate DNA strands of different lengths.

Prepare the gel

  1. Add 1 g of agarose powder to 50 mL of 1X TAE buffer.

  2. Heat the solution using the microwave until the agarose is completely dissolved.

    3. SAFETY AND USEFUL RECOMMANDATION
    When heating the mixture it is important to use an adaptable container. Putting metallic objects in the microwave is dangerous. It causes the formation of intense electric current. Keep in mind that the container might explode causing the deterioration of the microwave, the preparation and possible injuries to the people next to the device. To avoid this kind of situation the microwave is only used for short heating periods with the user keepin a eye on the preparation.

  3. Once the solution's temperature has decreased (enough to be able to touch the container), pour it into a gel cast (don't forget the comb).

Load the gel

  1. When the gel is solid, carefully remove the comb.

  2. Place the gel in the buffer tank and cover it with the 1X TAE buffer solution.

  3. Load each sample into the wells (don't forget the loading dye and the DNA ladder).

Run the gel

  1. Connect the generator.

  2. Chose the right voltage.

  3. Turn on the device.

    4. SAFETY AND USEFUL RECOMMANDATION
    The generator can deliver over 130V. The risk of electrocution for people is generally around 50V. That is why some recommendations must be followed. Before using the electrophoresis device, users should check if they have all parts of the device, because they are removable. They also have to check that each part corresponds to the right device. There are different kinds of tanks and parts are not compatible with each other. Using parts from another type of tank may lead to the loss of safety protections. We managed to assign a number to each corresponding part in order to know to which device they belong to. The generator must not be in presence of water and its vents must not be blocked. The generator has two main safety protections: a detector of migration buffer and a lid. The detection does not let the device generate electricity until the migration buffer has been put in the tank and reached the correct level. This is a protection both for the device and for users. Moreover, it prevents users to be surprised by an unexpected production of electricity during the manipulation. The second protection prevents the generator to produce electricity when no lid is put on the tank. With this, there is less risk of injuries and accidents, if it's correctly used.

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