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4. Disperse carefully to avoid air bubbles.
5. Pour into Petri dishes. Stack them one upon the other and Iet cool to
solidify.
6. Store the plates at 4 oc until used.
For transformation of JM103 with recombinant MB DNA (BRL-focus 1980)
• Recombinant M13 DNA
• A minimal agar plate (see above)
• 2 x YT agar plates (see above)
• 2 x YT top-agar (see above)
• E. coli JM103 grown on minimal medium
• 2 x YT medium (see above)
• 50 mM CaC}z (sterile and ice-cold)
• Glycerol sterilized by autoclaving (ice-cold)
• 100 mM IPTG (isopropyl-ß-D-thiogalactopyranoside), dissolved in sterile H20
• 2o/o X-gal solution (5-bromo-4-chloro-3-indolyl-ß-D-galactoside), dissolved in dimethylformamide (DMF)
• Shaker or shaking water bath
• Spectrophotometer
• Centrifuge
• Water bath
• Incubator
Procedure
Reagents
Solutions
Supplies
1. Streak JM103 cells on a minimal agar plate so that singlecolanies can be ln advance
obtained. Incubate the plate inverted at 37°C overnight.
2. Inoculate 10 ml of 2 x YT medium sterile with a single colony of JM103
grown on a minimal agar plate. Incubate the culture in a shakerat 37°C
and Iet the cells grow overnight.