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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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Fundamental
- Starting to work in the MEC group
- Maintaining lab notes
- How to mark your material
- Autoclaving
- Yeast and bacterial freezer stocks
- LGM
- Congresses
- Printable calendars
- Dept phone directory
- Antibiotics
- microscope
Cultivating the yeast Saccharomyces cerevisiae for any purpose
Teaching
The Yeast Pathway Kit
- The Yeast Pathway Kit
- pYPKa cloning protocol
- Transcription Unit cloning protocol
- standard_primers
- primer design
- in silico assembly of pYPKa_A_ATF1
- in silico assembly of pTA1_TDH3_ATF1_PGI1
- Plasmids, Promoters & Terminators sequence files.
Gel electrophoresis
- How many times can I reuse electrophoresis buffer?
- TAE
- fatval
- Turner
- Bachman
- Midori Green DNA loading buffer (LBx2wMidori)
- LiAc Borate
Plasmid DNA from E. coli
- Boiling miniprep
- CAVAN low cost plasmid miniprep protocol without kit (super snik)
- Merlin
- Plasmid midiprep protocol
DNA
- ethanol precipitation
- ligation
- restriction digestion
- ladders
- Extraction of chromosomal DNA with PCA (phenol:chloroform:isoamylalcohol)
- Rapid isolation of yeast total DNA
- Yeast genomic DNA extraction with LiOAc SDS
- Yeast DNA isolation with Lyticase or Zymolyase
- Plasmid rescue to E. coli with plasmid miniprep kit
- microwave yeast colony pcr
PCR
Transformation
E. coli
S. cerevisiae
Yeast cell (protein) extract from S. cerevisiae
RNA
Methods (miscellaneous)
E. coli strains
E. coli media
- LB
- LB-amp
- LB-kan
- LBpowder
- M9
- SOB (Super Optimal Broth)
- SOC (SOB with glucose)
- Super Broth
- Terrific Broth
- LB24
Yeast media
Stock solutions
- 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 ampicillin 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