Team:CIDEB-UANL Mexico/WetLab-Protocols
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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> |
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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 align="center"> <img src="https://static.igem.org/mediawiki/2013hs/2/2a/Imagen1.jpg" width="266px" height="265px" /> </p> | <p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/2/2a/Imagen1.jpg" width="266px" height="265px" /> </p> | ||
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<p> <b>Preparation for competent cells and their transformation</b></p> | <p> <b>Preparation for competent cells and their transformation</b></p> | ||
- | <p>This | + | <p>This i to prepare the plasmid in the bacterium with the thermal shock |
<br>Transformation Procedure: | <br>Transformation Procedure: | ||
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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> | ||
<p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/8/85/Imagen4.jpg" width="224px" height="227px" /> </p> | <p align="center"> <img src="https://static.igem.org/mediawiki/2013hs/8/85/Imagen4.jpg" width="224px" height="227px" /> </p> | ||
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<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: | ||
Revision as of 20:03, 21 June 2013
Wet-Lab
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Protocols
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General Rules:
Recommendations for Microbiology
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 i to prepare the plasmid in the bacterium with the thermal shock
Inoculation in Petri Dish and Test Tube Inoculate bacteria in dishes with agar and medium tubes with their antibiotics
Plasmid DNA miniprep This is to obtain the DNA plasmid of a colony
DNA quantification by UV spectrophotometer This is for quantify the plasmid DNA extracted
Plasmid DNA characterization This is for identify the piece we have
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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