Team:Colombia/Protocols
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Protocols
Contents |
Instructions for gel preparation:
- Weigh 0,3g of agarose.
- Add 30 mL of TAE 1x.
- 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.
- 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.
- When homogeneous, add 2 µL of SYBR SAFE DNA Gel Stain to the solution and mix well.
- 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.
- 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.
Note: For big gels use 0,7 g of agarose, 70 mL of TAE 1x and 3 µL of SYBR SAFE Gel Stain.
Instructions for LB medium preparation
LB liquid medium preparation (1 L)
- 10 g tryptone
- 10 g NaCl
- 5 g yeast extract
- Water
LB solid medium preparation (1 L)
- 15 g agar agar
- 10 g tryptone
- 10 g NaCl
- 5 g yeast extract
- Water
For selective medium, suplement with 20 ug/mL of antibiotic (kanamycin or chloramphenicol as appropiate).
COTENIDO 3
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COTENIDO 4
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COTENIDO 5
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COTENIDO 6
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COTENIDO 7
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