Difference between revisions of "Team:Manchester/Project/Protocols"
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Revision as of 23:08, 23 September 2016
Medium and Buffers
- Prepare the solution by mixing the ingredients stated above.
- Sterilize in an autoclave before using it to prepare the SOC medium.
- Prepare the solution by mixing the ingredients stated above.
- Sterilize in an autoclave.
*Source from 2015 iGEM Exeter [link = 2015.igem2015 .org/wiki/images/a/ab/Exeter_Glycerol_recipe.pdf]
For nucleic acid DNA/RNA separation
- TAE buffer (Tris-acetate-EDTA)
- TBE buffer (Tris-borate-EDTA)
- LB buffer (Lithium borate)
- Prepare the solution by mixing the ingredients stated above.
If not using pre-mixed LB agar powder, prepare the materials as below:
- In a 1L Erlenmeyer flask, swirl and mix the solution.
- Cover the top of the flask with a lid/aluminum foil and label with autoclave tape.
- Autoclave the liquid setting for 20 minutes or according to your autoclave's specifications.
- After removing the solution from the autoclave, allow the agar solution to cool to 55°C in an oven or water bath.
- When pouring the LB agar into plates, keep the bench area sterile by working near a flame or Bunsen burner. Alternatively, prepare the plates in a vacuum hood.
- Add the appropriate amount of desired antibiotic (refer the table below) to the solution and swirl to mix.
- Pour approximately 20mL of LB agar per 10cm polystyrene Petri dish.
- Place the lids on the plates and allow them to cool for until the agar is solidified.
- Label the bottom of plates with antibiotic and date before storing in plastic bags or sealed with Parafilm at 4°C.
Table source from New England Biolabs
Additional note:
- Antibiotic carbenicillin can be substituted for ampicillin in antibiotic selection plates [1].
