User:Jarle Pahr/Checklists
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Checklists for lab procedures:
Transformation:
- Supercompetent cells are available (remove from freezer 20 min before use, to avoid waiting for cells to thaw).
- Enough plates, with right antibiotic, are available. Remove from cold storage, place in incubator for pre-heating.
- Waterbath for heat shock is at right temperature (42 C).
Before heat shock: Place plates in incubator for pre-heating.
Restriction digest:
- Needed restriction enzymes are available.
- Thaw buffer
- Waterbath at 37 C is available. (or otherwise, thermomixer, or otherwise, shaking incubator)
Ligation:
- Ligation buffer is available.
- Ligase is available.
- Correct ligase is used (T4 DNA ligase)
- If possible include positive control to make sure the ligase is functional.
- If possible, include negative control of bacbone only/insert only, as appropriate.
Preparation of agar plates:
- Magnetic stirrers are available
- Antibiotic is available. Thaw antibiotic.
Preparing supercompetent cells:
- Centrifuge is cooled
- Spectrophotometer and cuvettes for measuring OD are available
- When using spectrophotometer: Correct wavelength is selected (600 nm)
- For snap freezing: Dry ice or liquid nitrogen is available.
- Freezer box is available
- Freezer box is marked.
Electrophoresis:
- GelDoc is operational
- After imaging: Save image as image file. Copy file(s) to personal area.
- Remove gel from imaging apparatus.
- Copy image to document, write observations.
Gel extraction:
- The correct band is cut.
End of day:
- No samples are left on bench (that shouldn't be there).