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This wiki contains protocols and other information mostly 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
- Congresses
- Printable calendars
- Dept phone directory
- EGB25
- EGB24
- EGB23
- GBM23
- GMB21
- LIB20
- Purifying and testing recombinant Taq DNA polymerase
- TP26 Simply Cloning
- 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.
- How many times can I reuse electrophoresis buffer?
- TAE
- fatval
- Turner
- Bachman
- Midori Green DNA loading buffer (LBx2wMidori)
- LiAc Borate
- Boiling miniprep
- CAVAN low cost plasmid miniprep protocol without kit (super snik)
- Merlin
- Plasmid midiprep protocol
- Plasmid rescue from yeast to E. coli with plasmid miniprep kit
Colony PCR from yeast is a notoriously fickle protocol which can be seen by the many variations available online. It seems many users have to tweak their protocols for them to work. The protocols below are listed in order of difficulty. Test the protocol with a positive and negative control if you can.
- microwave yeast colony pcr
- Yeast genomic DNA extraction with LiOAc SDS
- Yeast DNA isolation with Lyticase or Zymolyase
- Rapid isolation of yeast total DNA
- Extraction of chromosomal DNA with PCA (phenol:chloroform:isoamylalcohol)
- LB
- LB-amp
- LB-kan
- LBpowder
- M9
- SOB (Super Optimal Broth)
- SOC (SOB with glucose)
- Super Broth
- Terrific Broth
- LB24
- 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
- Antibiotics