TFB1

Chemicals Mass (g) RbCl 3.02965 MnCl2 1.56996 Potassium Acetate 0.74392 CaCl2 0.27806 Glycerol 37.5463	TFB2 Chemicals Mass (g) MOPS 0.53398 RbCl 0.30225 CaCl2 2.88320 Glycerol 37.5457

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Making Antibiotic Stocks

02/1/13

Antibiotics are used to isolate organisms into which plasmids containing antibiotic resistance genes have been transformed.

Make antibiotic plates with the following specs.

Antibiotic	Concentration (µg/mL)	Color Code
Ampicillin	100	Orange
Chloramphenicol	35	Green
Kanamycin	50	Red
Tetracycline	15	Yellow

Stocks should be made at 1000x concentration, so that making 1L of plate medium will require only 1mL of stock antibiotic solution.

Stocks of antibiotics are made at the following concentrations

Ampicillin	100 mg/mL
Chloramphenicol	35 mg/mL
Kanamycin	30 mg/mL
Tetracycline	15 mg/mL

50mL of stock were prepared and split into several 15mL tubes

Stocks should be stored in the refrigerator at 4°C

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01/25/13

Restreaked TOP10 cells on LB plates for isolation of single colonies.

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01/23/13

Sealed TOP10 E. coli cells on LB agar plate. Stored in 37°C incubator

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01/18/13

Poured LB agar plates

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01/16/13

Making Agar Plates

LB agar is used to grow our stocks of Escherichia coli

• 10g Bacto Tryptone
• 5g Yeast Extract
• 10g Sodium Chloride (NaCl)
• 15g Agarose

Mix Components in 1L of dH₂O until dissolved. Spilt 1000mL solution into two flask. Cap flask with aluminum to prevent spilling of solution. Autoclave on slow exhaust for 20 minutes. Keep liquid in 65°C water bath to prevent setting until 18-Jan.

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