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This wiki contains protocols and other information of interest to the MEC group at CBMA and people working in the LGM lab.
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- Starting to work in the MEC group
- Maintaining lab notes
- How to mark your material
- Autoclaving
- Yeast and bacterial freezer stocks
- LGM
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- Antibiotics
Chromosomal DNA / Total DNA / Genomic DNA / gDNA / plasmid DNA / pDNA preparation from S. cerevisiae
This protocol is perhaps quick but also dirty. There should be considerable amounts of phenol in the final preparation. Do not use for E. coli transformation, the phenol kills the E.coli.
This protocol is similar to the one above, but incorporates an ethanol precipitation at the end. This has been extensively used and usually works well. At one point, PCR was not possible using the undiluted DNA as template, but PCR products appeared from x10 dilutions. Possibly due to oxidized phenol.
This protocol is one of the most cited in literature. This is an indication that it works well.
This protocol does not include glass beads, so it should yield higher molecular weight DNA fragments.
This protocol allow isolation of enough plasmid DNA to transform E. coli. The DNA can also be used for PCR.
- bradford
- disintegration protocol
- LiAc Borate agarose gel electrophoresis
- pYPKa cloning protocol
- Transcription Unit cloning protocol
- Pathway assembly with the Yeast Pathway Kit
- sequencing
- Taq purification
- yeast colony pcr
- Yeast cell fixation with formaldehyde
- 2xPCR mastermix
- Agar 40 g/L (2x)
- 40% w/v Glucose stock solution for media
- 40% w/v Sucrose stock solution for media
- 25% w/v NaCl stock solution for media
- 3.5M KCl stock solution.
- 2.5M MgCl2 stock solution.
- 2M MgSO4 stock solution.
- 0.5 M EDTA stock solution, pH 8
- 1.0 M TrisHCl, pH 8
- 1M IPTG stock solution
- 200 mg/mL amp icillin solution (x1000)
- 200 mg/mL Geneticin solution (x1000)
- 12.5 mg/mL Tetracycline solution (x1000)
- 10 mM dNTPs solution
- 5x Ligase buffer
- DNA DNA loading-buffer with FICOLL and two dyes.
- PBS