Invitrogen Bac to Bac Expression System

Appendix
Recipes
Antibiotic Stock
Solutions
Antibiotics can be ordered in either dry powdered form or as a stabilized, sterile,
premixed solution. Store these solutions according to the manufacturer’s
recommendations. For the antibiotics below, prepare and store the stock
solutions as directed:
Antibiotic Stock Solution Concentration Storage
Ampicillin 50 mg/ml in water; filter-sterilize -20°C, protected from light
Kanamycin 10 mg/ml in water; filter-sterilize -20°C, protected from light
Tetracycline* 10 mg/ml in water; filter-sterilize -20°C, protected from light
Gentamicin 7 mg/ml in water; filter-sterilize -20°C, protected from light
*If you are using tetracycline from Invitrogen (Catalog no. Q100-19), prepare in water. If you are
using the free-base form of tetracycline, prepare in 100% ethanol.
IPTG
Follow the procedure below to prepare a 200 mg/ml stock solution of IPTG.
1. Dissolve 2 g of IPTG in 8 ml of sterile water.
2. Adjust the volume of the solution to 10 ml with sterile water.
3. Filter-sterilize through a 0.22 micron filter.
4. Dispense the stock solution into 1 ml aliquots.
5. Store at -20°C.
Bluo-gal
Follow the guidelines below to prepare a 20 mg/ml stock solution of Bluo-gal.
• Dissolve the Bluo-gal in dimethylformamide or dimethyl sulfoxide (DMSO)
to make a 20 mg/ml stock solution. Use a glass or polypropylene tube.
Important: Exercise caution when working with dimethylformamide.
Dispense solutions in a vented chemical hood only.
• Do not filter the stock solution.
• Store at -20°C protected from light.
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