Difference between revisions of "Team:Nagahama/Experiments"
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{{Nagahama}} | {{Nagahama}} | ||
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+ | == '''Protocols''' == | ||
+ | |||
+ | |||
+ | |||
+ | <br><br> | ||
+ | |||
+ | == Agarose gel == | ||
+ | |||
+ | <br><br> | ||
+ | |||
+ | <li>Weigh 0,3 g of agarose.</li> | ||
+ | <li>Add 30 mL of TAE 1X.</li> | ||
+ | <li>Heat up until the solution is homogeneous, avoiding boiling. If it boils, move away from the heat until it “calms down” and put it back on the heat until the agarose is completely dissolved.</li> | ||
+ | <li>While heating, prepare the bed in which the gel will polymerize. Make sure that it is well balanced and tight, and that the “comb” is well placed.</li> | ||
+ | <li>When homogeneous, add 2 µL of SYBR SAFE DNA Gel Stain to the solution and mix well.</li> | ||
+ | <li>Pour the solution into the bed and clear all its bubbles with a tip. Put a piece of paper on top of it and let it polymerize.</li> | ||
+ | <li>Mix the samples with loading dye in a 5:1 ratio. Put the samples into the wells, as well as 4 µL of molecular weight marker into the first well.</li> | ||
+ | </ol> | ||
+ | <b>Note</b>: For big gels use 0,7 g of agarose, 70 mL of TAE 1x and 3 µL of SYBR SAFE Gel Stain. | ||
+ | |||
+ | |||
+ | |||
+ | == LB medium (1L liquid) == | ||
+ | <br><br> | ||
+ | |||
+ | |||
+ | <ul> | ||
+ | <li>10 g tryptone</li> | ||
+ | <li>10 g NaCl</li> | ||
+ | <li>5 g yeast extract</li> | ||
+ | <li>Water</li> | ||
+ | </ul> | ||
+ | |||
+ | <br><br><br> | ||
+ | |||
+ | == LB medium (solid, 1L = 50 dishes) == | ||
+ | <br><br> | ||
+ | <p align="justify"> | ||
+ | <ul> | ||
+ | <li>15 g agar agar</li> | ||
+ | <li>10 g tryptone</li> | ||
+ | <li>10 g NaCl</li> | ||
+ | <li>5 g yeast extract</li> | ||
+ | <li>Water</li> | ||
+ | </ul> | ||
+ | </p> | ||
+ | For selective medium, suplement with antibiotic as appropiate (kanamycin 50 µg/ mL and 100 µg/mL for chloramphenicol or ampicillin ). | ||
+ | |||
+ | |||
+ | <br><br><br> | ||
+ | |||
+ | == Genome extraction == | ||
+ | <br><br> | ||
+ | |||
+ | For bacterial genome extraction we used Easy DNA Kit, Invitrogen according to <a href="https://2014.igem.org/File:Easy-DNA_Kit.pdf" target="_blank">manufacturer's instructions</a>. | ||
+ | |||
+ | <br><br><br> | ||
+ | |||
+ | == Plasmids extraction == | ||
+ | |||
+ | <br><br> | ||
+ | For bacterial plasmid extraction we used GenElute Plasmid Miniprep Kit, Sigma Aldrich according to <a href="https://2014.igem.org/File:Miniprep.pdf" target="_blank">manufacturer's instructions</a>. | ||
+ | |||
+ | |||
+ | <br><br><br> | ||
+ | > | ||
<html> | <html> | ||
Revision as of 06:22, 30 May 2015
Contents
Protocols
Agarose gel
</ol> Note: For big gels use 0,7 g of agarose, 70 mL of TAE 1x and 3 µL of SYBR SAFE Gel Stain.
LB medium (1L liquid)
- 10 g tryptone
- 10 g NaCl
- 5 g yeast extract
- Water
LB medium (solid, 1L = 50 dishes)
- 15 g agar agar
- 10 g tryptone
- 10 g NaCl
- 5 g yeast extract
- Water
For selective medium, suplement with antibiotic as appropiate (kanamycin 50 µg/ mL and 100 µg/mL for chloramphenicol or ampicillin ).
Genome extraction
For bacterial genome extraction we used Easy DNA Kit, Invitrogen according to <a href="https://2014.igem.org/File:Easy-DNA_Kit.pdf" target="_blank">manufacturer's instructions</a>.
Plasmids extraction
For bacterial plasmid extraction we used GenElute Plasmid Miniprep Kit, Sigma Aldrich according to <a href="https://2014.igem.org/File:Miniprep.pdf" target="_blank">manufacturer's instructions</a>.
>
Experiments & Protocols
Describe the experiments, research and protocols you used in your iGEM project.
What should this page contain?
- Protocols
- Experiments
- Documentation of the development of your project