- 1 How do you add antibiotics to agar?
- 2 How is antibiotic stock solution prepared?
- 3 How do you prepare media?
- 4 Why do we add antibiotics to culture media?
- 5 How much antibiotics add to agar plates?
- 6 How is antibiotic dilution calculated?
- 7 How do you make 100 micrograms per ml solution?
- 8 Is chloramphenicol an antibiotic?
- 9 How do you prepare and sterilize nutrient agar?
- 10 What is the purpose of media preparation?
- 11 What bacteria does not grow on blood agar?
- 12 What is the purpose of the antibiotics in the agar plate?
- 13 Why is penicillin used in cell culture?
- 14 What does the Zone of Inhibition tell us?
How do you add antibiotics to agar?
Add antibiotic stock (200 µl for 200 ml) to the liquid LB- agar and slowly mix. Bunsen burner flame. Close the lid after filling the plate. Let the agar solidify for ~1 hour on the bench.
How is antibiotic stock solution prepared?
Preparation of 80ml stock solution
- Tetracycline is kept in the 4C fridge in 68-564D.
- Weigh 400mg of tetracyline HCL into a small weigh boat.
- Dilute 95% Ethanol to 70% using milliQ water.
- Add 80ml of 70% Ethanol to a 250ml bottle.
- Add the tetracycline HCL to the ethanol.
How do you prepare media?
It is really very simple to make complex media these days:
- rehydrate the powder form of the medium.
- stir and boil the agar medium to get the agar powder dissolved (if making an agar medium rather than a broth medium )
- distribute the medium into tubes.
- autoclave to sterilize the tube media.
Why do we add antibiotics to culture media?
Use of antibiotics in cell culture minimizes the loss of valuable cells, reagents, time and efforts due to contamination. Maintenance of aseptic conditions and techniques is vital to a research laboratory that handles cell/tissue culture.
How much antibiotics add to agar plates?
Prepare carbenicillin to a concentration of 1 mg/mL – 4 mg/mL in LB medium. The concentration of antibiotic required for effective over- agar selection has been empirically determined.
How is antibiotic dilution calculated?
The dilution factor is: (5 mM) / (50 µM) = (5000 µM) / 50 µM) = 100 so you need to dilute: (3 ml) / 100 = (3000 µl) / 100 = 30 µl of the stock solution to a final volume of 3 ml.
How do you make 100 micrograms per ml solution?
Take 1ml of the stock solution (covert(100ug/ ml ) and add 9mls of water to produce a100ug/10ml solution. 1ml of this will give 10ug/ ml.
Is chloramphenicol an antibiotic?
Chloramphenicol is an antibiotic. It’s mainly used to treat eye infections (such as conjunctivitis) and sometimes ear infections. Chloramphenicol comes as eye drops or eye ointment.
How do you prepare and sterilize nutrient agar?
How to prepare nutrient agar?
- Suspend 28g of nutrient agar powder (CM0003B) in 1L of distilled water.
- Mix and dissolve them completely.
- Sterilize by autoclaving at 121°C for 15 minutes.
- Pour the liquid into the petri dish and wait for the medium to solidify.
What is the purpose of media preparation?
To become familiar with the necessary nutritional and environmental factors for culturing microorganisms in the laboratory. To understand the decontamination or sterilization process using an autoclave. To learn the procedures used in preparing media needed for culturing microorganisms.
What bacteria does not grow on blood agar?
Fastidious organisms, such as streptococci, do not grow well on ordinary growth media but grow on blood agar.
What is the purpose of the antibiotics in the agar plate?
The addition of an antibiotic to this gel allows for the selection of only those bacteria with resistance to that antibiotic – usually conferred by a plasmid carrying the antibiotic resistance gene. The following protocol will allow you to make your own LB/ agar plates with your antibiotic of interest.
Why is penicillin used in cell culture?
The antibiotics penicillin and streptomycin are used to prevent bacterial contamination of cell cultures due to their effective combined action against gram-positive and gram-negative bacteria.
What does the Zone of Inhibition tell us?
The Zone of inhibition is a circular area around the spot of the antibiotic in which the bacteria colonies do not grow. The zone of inhibition can be used to measure the susceptibility of the bacteria to wards the antibiotic.