Team:CIDEB-UANL Mexico/WetLab-Protocols
From 2013hs.igem.org
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- | <p> <b>General Rules</b> | + | <p> <b>General Rules:</b> |
- | <br>1. - | + | <br>1. - Always use your lab coat |
- | <br>2. - | + | <br>2. - Always pick up the used material |
<br>3. - Never take any material to your mouth while you’re working with reactants | <br>3. - Never take any material to your mouth while you’re working with reactants | ||
- | <br>4. - Keep | + | <br>4. - Keep your working area clean |
- | <br>5. - | + | <br>5. - All non-reusable material must be placed in the correct trash can |
- | <br>6. - Check gas and water | + | <br>6. - Check for oppen gas and water valves, depending on your work |
<br>7. - Wash your hands before and after every session | <br>7. - Wash your hands before and after every session | ||
- | <br>8. - Always wear gloves | + | <br>8. - Always wear gloves when working with Ethidium bromide |
- | <br>9 | + | <br>9. - Every question, accident, etc. tell the instructor</p> |
- | + | ||
<p><b>Recommendations for Microbiology</b> | <p><b>Recommendations for Microbiology</b> | ||
- | <br> | + | <br>10. - Sterilize with heat any crop material before and after using it. |
- | <br> | + | <br>11. - Don’t talk while you are working with sterilized material |
- | <br> | + | <br>12. - When you start and finish, sterilize the surface with 70% alcohol |
- | <br> | + | <br>13. - Al testing tubes, must be vertical |
- | <br> | + | <br>14. - Never deposit a living cultivation in the sink |
- | <br> | + | <br>15. - Sterilize every used material</p></td> |
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+ | <p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/d/da/Imagen2.jpg" width="202px" height="203px" /> </p> | ||
+ | </td> | ||
+ | </tr></table> | ||
+ | <table style="background-color: #FFFFFF;" width="100%" id="texto"> | ||
+ | <tr> | ||
+ | <td> | ||
<p> <b>Laboratory Protocol </b></p> | <p> <b>Laboratory Protocol </b></p> | ||
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<p> This step is for searching and obtaining the desired piece. | <p> This step is for searching and obtaining the desired piece. | ||
<br>Procedure: | <br>Procedure: | ||
- | <br>1.- Search in the Parts Registry | + | <br>1.- Search for the BioBrick in the Parts Registry |
<br>2.- Drill the plate, and add 10ul of mQ water | <br>2.- Drill the plate, and add 10ul of mQ water | ||
- | <br>3.- Mix very well, and take the | + | <br>3.- Mix very well, and take the resusupended DNA to an Eppendorf tube </p> |
- | + | ||
<p> <b>Preparation for competent cells and their transformation</b></p> | <p> <b>Preparation for competent cells and their transformation</b></p> | ||
- | <p>This is | + | <p>This is to prepare the plasmid in the bacterium with the thermal shock |
<br>Transformation Procedure: | <br>Transformation Procedure: | ||
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<br>4.- Dive the Eppendorfs with 42°C water for 1 minute | <br>4.- Dive the Eppendorfs with 42°C water for 1 minute | ||
<br>5.- Take it to ice for 2 minutes | <br>5.- Take it to ice for 2 minutes | ||
- | <br>6.- Add LB and incubate them for 20 min, then put them into petri dish at 37°C all the night. </p> | + | <br>6.- Add LB and incubate them for 20 min, then put them into petri dish at 37°C all the night. </p></td> |
+ | <td style="padding:12px;"> | ||
+ | <p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/2/2a/Imagen1.jpg" width="266px" height="265px" /> </p> | ||
+ | </td></tr></table> | ||
+ | <table style="background-color: #FFFFFF;" width="100%" id="texto"> | ||
+ | <tr> | ||
+ | <td> | ||
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+ | <table style="background-color: #FFFFFF;" width="100%" id="texto"> | ||
+ | <tr> | ||
+ | <td> | ||
<p><b> Inoculation in Petri Dish and Test Tube </b></p> | <p><b> Inoculation in Petri Dish and Test Tube </b></p> | ||
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<br><b>For planting</b> | <br><b>For planting</b> | ||
<br>1.- Take the colonies in a test tube with LB and the antibiotic | <br>1.- Take the colonies in a test tube with LB and the antibiotic | ||
- | <br>2.- Take the handle sterilized and spread the colonies in | + | <br>2.- Take the handle sterilized and spread the colonies in a Petri dish. |
<br>3.- Incubate for 37°C FOR 16 hours | <br>3.- Incubate for 37°C FOR 16 hours | ||
<br>Reseeding | <br>Reseeding | ||
- | <br>1.- Add the antibiotic to a test tube | + | <br>1.- Add the antibiotic to a test tube previously half filled with Lb medium |
- | <br>2.- | + | <br>2.- With a handle take a colony |
<br>3.- Then agitate the handle in the Tube, finally incubate at 37°C </p> | <br>3.- Then agitate the handle in the Tube, finally incubate at 37°C </p> | ||
- | + | </td> | |
- | + | <td><p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/8/85/Imagen4.jpg" width="224px" height="227px" /> </p></td></tr></table> | |
+ | <p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/b/b4/Imagen3.jpg" width="500px" height="200px" /> </p> | ||
+ | <table style="background-color: #FFFFFF;" width="100%" id="texto"> | ||
+ | <tr> | ||
+ | <td> | ||
<p><b>Plasmid DNA miniprep</p></b> | <p><b>Plasmid DNA miniprep</p></b> | ||
- | <p>This is | + | <p>This is to obtain the DNA plasmid of a colony |
<br>Procedure: | <br>Procedure: | ||
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<br>7.- Centrifuge again and throw the liquid | <br>7.- Centrifuge again and throw the liquid | ||
<br>8.- Let the precipitate incubate at 37°C, then add 200ul of mQ water with RNase and mix it | <br>8.- Let the precipitate incubate at 37°C, then add 200ul of mQ water with RNase and mix it | ||
- | <br>9.- Check in the gel or save it at 4°C </p> | + | <br>9.- Check in the gel or save it at 4°C </p></td> |
- | + | <td><p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/e/e6/Imagen_1.jpg" width="132px" height="285px" /> </p></td></tr></table> | |
- | + | <table style="background-color: #FFFFFF;" width="100%" id="texto"> | |
+ | <tr> | ||
+ | <td> | ||
<p><b> DNA quantification by UV spectrophotometer</b> </p> | <p><b> DNA quantification by UV spectrophotometer</b> </p> | ||
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<br>4.- Pass the DNA to the spectrophotometer | <br>4.- Pass the DNA to the spectrophotometer | ||
<br>5.- Select the option (DNA or RNA) | <br>5.- Select the option (DNA or RNA) | ||
- | <br>6.- Write the lecture of the spectro at 260, 280 and 320, relation 260/280 and the concentration.</p> | + | <br>6.- Write the lecture of the spectro at 260, 280 and 320, relation 260/280 and the concentration.</p></td></tr></table> |
- | + | ||
+ | <table style="background-color: #FFFFFF;" width="100%" id="texto"> | ||
+ | <tr> | ||
+ | <td> | ||
<p> <b>Plasmid DNA characterization </b></p> | <p> <b>Plasmid DNA characterization </b></p> | ||
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<br>4.- Incubate the reactions at 37°C for 1 hour | <br>4.- Incubate the reactions at 37°C for 1 hour | ||
<br>5.- Use the gel for check the result </p> | <br>5.- Use the gel for check the result </p> | ||
- | + | </td> | |
- | + | <td><p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/a/af/Imagen6.jpg" width="282px" height="232px" /> </p></td></tr></table> | |
<p><b>BioBrick parts assembly</b></p> | <p><b>BioBrick parts assembly</b></p> | ||
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<br>5.- Incubate the reactions at 25°C</p> | <br>5.- Incubate the reactions at 25°C</p> | ||
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</td> | </td> | ||
</tr> | </tr> |
Latest revision as of 01:14, 22 June 2013
Wet-Lab
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Protocols
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General Rules:
Recommendations for Microbiology
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Laboratory Protocol Handling of lyophilized DNA’s plates. This step is for searching and obtaining the desired piece.
Preparation for competent cells and their transformation This is to prepare the plasmid in the bacterium with the thermal shock
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BioBrick parts assembly Procedure:
Ligation of genetic parts This step is for paste the desired piece in the bacterium we are going to use.
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CIDEB UANL Team. Centro de Investigación y Desarrollo de Educación Bilingüe |
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