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.