How much kanamycin do you put on a LB plate?

How much kanamycin do you put on a LB plate?

Autoclave 1 L of LB agar. Cool to 55°C. Add 1 mL of the kanamycin stock (i.e., 50 mg).

How do you make LB agar with kanamycin?

1. Prepare LB Agar plates: add 3.7 g LB Agar into 100 mL H2O, autoclave, cool agar to 42º C, add antibiotics (ampicillin (100 mg/L) and kanamycin (100 mg/L)), pour liquid agar into 4 plates, and let agar harden for 1-2 hours. 2. Thaw competent cells (DH5α) slowly on ice.

Why is kanamycin added to the agar?

Kanamycin is used in molecular biology as a selective agent most commonly to isolate bacteria (e.g., E. Bacteria that have been transformed with a plasmid containing the kanamycin resistance gene are plated on kanamycin (50-100 ug/ml) containing agar plates or are grown in media containing kanamycin (50-100 ug/ml).

What is the working concentration of kanamycin?

50 mg/mL
Antibiotic Concentrations

Antibiotic Recommended Stock Concentration Recommended Working Concentration
Coumermycin 25 mg/mL (dissolve in DMSO) 25 µg/mL
Gentamycin 10 mg/mL 10 µg/mL
Kanamycin 50 mg/mL 50 µg/mL
Spectinomycin 50 mg/mL 50 µg/mL

How much chloramphenicol do I add to LB broth?

Chloramphenicol – The frozen stock solutions of chloramphenicol are at 25mg/ml in 100% ethanol and are marked with purple. The final concentration for LB liquid culture is 25mg/ml. To obtain this in 100ml of LB, add 100ul stock solution.

Is E coli sensitive to kanamycin?

coli strains isolated from well neonates, well pediatric outpatients, and ill neonates were determined, and the incidence of antibiotic-resistant strains was correlated with epidemiologic data. Ninety-five per cent of these isolates were susceptible to kanamycin and 98 per cent to gentamicin.

Can you vortex kanamycin?

It is light sensitive. Weigh 800mg of kanamycin into a small weigh boat. Add 80ml of milliQ to a 250ml bottle. Mix/vortex so all the kanamycin goes into solution.

How much volume of ampicillin stock 100 mg/ml will be added in 100 ml of LB broth so that final concentration would be 100 μg μl?

Ampicillin – The frozen stock solutions of ampicillin are at 50mg/ml and 100mg/ml in H2O, and are marked with a red sticker. The final concentration for LB liquid culture is 50ul/ml. To obtain this in 100ml (the amount in each LB bottle), add 100ul stock solution.