Richard Lab:Microorganisms & Media

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Prokaryotic

E. coli NEB 5a

• A DH5-alpha derivitive supplied by New England Biolabs
• Used for general cloning including Bgal selection.
• Electrocompetent cells are kept in the -80°C Freezer in Dr. Demirci's lab.
• Ask Mike

E. coli DB3.1

• These clones are subject to ccdb lethality
• Plasmids with the ccdb gene surrounding the MCS are used to select for inserts
  • DB3.1 cells with non insert plasmids will die
• Ask Mike

Lactobacillus plantarum PA-28

• Commercially available silage inoculant
• Found in "Biomax 5" by CHR Hansen biosystems
• Ask Mike

Lactobacillus plantarum K-270

• Commercially available silage inoculant
• Found in "Biomax 5" by CHR Hansen biosystems
• Ask Mike

Lactobacillus plantarum LP-401

• Commercially available silage inoculant
• Found in "Biomax MP" by CHR Hansen biosystems
• Ask Mike

Lactobacillus plantarum LP-1

• Commercially available silage inoculant
• Found in "Biomax MP" by CHR Hansen biosystems
• Ask Mike

Lactobacillus plantarum MD007

• is a knockout of the alanine racemase gene
• cannot grow on MRS media without supplemented D-alanine
• used for promoter probing with the pNZ7120 promoter probe vector
• Bron, P.A., S. M. Hoffer, I. I. Van Swam, W. M. De Vos and M. Kleerebezem. 2003. Selection and Characterization of Conditionally Active Promoters in Lactobacillus plantarum, Using Alanine Racemase as a Promoter Probe. Applied and Environmental Microbiology 70 (1): 310–317
• Ask Mike

Lactococcus lactis MG1363

• Contains pNZ7120 promoter probe vector
• Can be selected for based on ampicillin resistance
• grow on M17 media supplemented with 0.5% glucose
• Bron, P.A., S. M. Hoffer, I. I. Van Swam, W. M. De Vos and M. Kleerebezem. 2003. Selection and Characterization of Conditionally Active Promoters in Lactobacillus plantarum, Using Alanine Racemase as a Promoter Probe. Applied and Environmental Microbiology 70 (1): 310–317
• Ask Mike

Pedicoccus pentosaceus PC-3

• Commercially available silage inoculant
• Found in "Biomax MP" by CHR Hansen biosystems
• Ask Mike

Eukaryotic

Saccharomyces cerevisiae NRRL Y-2034 (ATCC 24702)

• Obtained from USDA-ARS Culture Collection
• Culture maintenance protocol
• Ask Megan

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Media

MRS Medium

• 10g/L Peptone
• 5g/L Yeast Extract
• 5g/L Sodium Acetate
• 2g/L Dipotassium Phosphate
• 2g/l Ammonium Citrate
• 1g/L Tween-80 (1ml polysorbate-80)
• 0.1g/L Magnesium Sulfate
• 0.05g/L Manganese Sulfate (137μL 38% Manganous Sulfate solution)
• Carbon source:
  • For Full strength medium add 20g/L dextrose and 10g/L beef extract.
  • For other purposes as little as 1.8g/L (10mM) dextrose can be used.
    • If adjusting the pH with malate or arginine use 2.5g/L dextrose.