User:Jarle Pahr/Checklists

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.

Measuring OD:

• Instrument is set to correct wavelength (typically 600 nm)
• Cultures are re-placed in incubator immediately after sampling.
• Enough liquid in cuvette
• Cuvette is inserted in correct orientation

Use of instruments, general:

• After end of run/experiment, copy data to personal area.
• Print paper of copy of raw or processed data, place in ring binder or lab journal.

Receiving plasmid samples for use:

• As soon as possible, verify the sequence (entirely, or critical parts only) for yourself
• Make a new glycerol stock for your personal use.

End of day:

• No samples are left on bench (that shouldn't be there).