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 Stock, concPreparationVolumes, TubesStorage
 10× T4 ligase bufferThawed, precipitates resuspended with vortexing and/or brief 37° incubation.50 µL aliquots-20°, rack 1.5
 10 mM dNTPs?20 µL aliquots-20°, rack 1.5
 Antibiotics
 chl25 (mg/mL)Ethanol, -20°; stable for ≥1 year.15 mL-20°, freezer 1 door
 amp10040% ethanol, -20°; seems to be stable for ≥6 mo for agar15 mL-20°, freezer 1 door
 kan50Water, 0°; stable for ≥1 year.15 mL-20°, freezer 0 door
 spt50Water, 0°; stable for ≥1 year.15 mL-20°, freezer 0 door

Antibiotics

AntibioticTarget Stock ConcentrationSolventsNotesRich Medium
  Abbr*Relativem/V  c, Multi-Copy; Rangec, Single-Copy
Chloramphenicolribosomechl1000×

25 mg/mL

(formerly 34 mg/mL)

100% Ethanol 25 or 34 mg/L   (3)≥25–340 mg/L   (5)10 mg/L (1)
Ampicillin·Napeptidoglycanamp1000×100 mg/mL40% ethanol or water

40% EtOH keeps it liquid at -20°C.
Substitutes carbenicillin.

100 mg/L   (3) 50 mg/L   (1)
Kanamycin B·SO4 ribosomekan1000×50 mg/mL35% DMSO or Water35% DMSO keeps it liquid at -20°C.Insoluble in 40% ethanol.
Precipitates from 35% DMSO.
50 mg/L   (3)≥50–500 mg/L   (5)20 mg/L   (1)
Spectinomycin·2HCl·5H2O ribosomespt1000×50 mg/mLWaterto be confirmed:
Precipitates from 35% DMSO keeps it liquid at -20°C.
50 mg/L   (3)≥50–500 mg/L   (5)20 mg/L   (1)
Streptomycin·SO4ribosomestr1000×100 mg/mLWater    
TrimethoprimFolA (DHFR)tmp1000×15 mg/mLDMSO or 1% acetic acidInsoluble in water or ethanol.15 mg/L? 10 mg/L (2)
Apramycin·SO4ribosomeapr1000×??Water, presumably ? ?
Gentamicin·SO4ribosomegen1000×10 mg/mL?Water or ? 10 mg/L 10 mg/L   (1)
Tetracycline·HClribosometet1000×10 mg/mLWater or ? 10 mg/L   (3) 15 mg/L   (1)
Hygromycin B?ribosomehyg1000×200 mg/mLWater 200 mg/L   (3)  


References

* Follow American Society of Microbiology abbreviation guidelines when possible

  1. Poteete et al. Biotechniques  (2006)  
  2. Kittleson et al.J Biol Eng (2011)
  3. Addgene
  4. Qiagen
  5. Sara Molinari, unpublished data, p15A with cat, aphA1aadA.