What is Zeocin used for?
Usage. Zeocin and other related chemicals in the bleomycin family of compounds are primarily used in molecular biology as an antibiotic, especially for the selection of eukaryotic cell lines when used in conjunction with vectors containing a selectable marker for zeocin resistance.
Is Zeocin an antibiotic?
Gibco™ Zeocin™ Selection Antibiotic is a formulation of phleomycin D1, a glycopeptide antibiotic produced by Streptomyces verticillus. Zeocin™ Selection Antibiotic acts by binding to and cleaving DNA, leading to cell death.
How long does Zeocin take to work?
In general, it takes 2-6 weeks to generate foci with Zeocin™, depending on the cell line.
Does Zeocin expire?
For Zeocin® powder, once resuspended at 100 mg/ml, it can be stored at 4°C for 12 months or -20 °C for 18 months.
Is Zeocin light sensitive?
Zeocin™ is light sensitive. Store the antibiotic and plates or medium containing the antibiotic in the dark. Reduce the salt in bacterial medium and adjust the pH to 7.5 to keep Zeocin™ active as high ionic strength and acidity or basicity inhibit Zeocin™ activity.
How do you make Zeocin stock solution?
Making the stock solution: Weigh out 1 g of ZeocinTM (Phleomycin, Gold Biotechnology, Catalog #Z-200 [CAS 11006-33-0, MW: 1526.5]). 2. Add 10 ml of sterile H2O, or Hepes Buffer (5 g/L, pH 7.25). Dissolve completely by mixing or vortexing.
How does chloramphenicol work?
Chloramphenicol stops bacterial growth by binding to the bacterial ribosome (blocking peptidyl transferase) and inhibiting protein synthesis. Chloramphenicol is lipid-soluble, allowing it to diffuse through the bacterial cell membrane.
What is killing curve?
A kill curve is a dose-response experiment where the cells are subjected to increasing amounts of antibiotic to determine the minimum antibiotic concentration needed to kill all the cells over the course of one week.
Can I use neomycin instead of G418?
Bottom line: Use G418 for single-selection experiments on eukaryotic cells. Use neomycin or kanamycin for single-selection experiments on prokaryotic cells where you are selecting for neomycin resistance genes. Kanamycin should be used over neomycin if elimination of Mycoplasma species from culture is necessary.
How do you make antibiotic solution?
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 select antibiotics?
Determining the Optimal Selection Antibiotic Concentration
- Plate cells in 0.5 ml complete growth medium per well in a 24-well tissue culture plate one day prior to introducing antibiotic selection.
- Add increasing amounts of the appropriate antibiotic such as G418 to duplicate wells of cells plated in complete media.
What is a checkerboard assay?
In the checkerboard assay, two antimicrobics are tested in double serial dilutions, and the concentration of each drug is tested both alone and in combination. Thus, it is possible to determine the effect of the individual drug, but above all, the effect produced by their combination.
What is the difference between G418 and neomycin?
Neomycin and G418 both will target cells which do not contain these genes and they will be eliminated. In general, neomycin is used in experiments on prokaryotic cells, while G418 is used in eukaryotic experiments.
How do you dilute an antibiotic solution?
Working dilution: 1/1000, so you have to put 500 µL stock solution in 500 mL of your medium, for example. Your final working concentration is now 100 µg/mL. Working dilution: 1/100, so you have to put 5 mL stock solution in 500 mL of your medium, for example.
Why use phleomycin instead of zeocin?
Use of phleomycin is recommended for cells poorly sensitive to Zeocin™, i.e. filamentous fungi and some yeasts. Phleomycin resistance is conferred by the Sh ble gene from Streptoalloteichus hindustanus which encodes a protein that binds to phleomycin, inhibiting its DNA cleavage activity.
How important is zeocin™ for selection in mammalian cells?
For selection in mammalian cells, physiological ionic strength and pH are much more important for cell growth, so more Zeocin™ may be needed for selection relative to yeast or bacteria.
What type of drug is zeocin?
Zeocin. Zeocin is a formulation of phleomycin D1, a glycopeptide antibiotic and one of the phleomycins from Streptomyces verticillus belonging to the bleomycin family of antibiotics. It is a broad-spectrum antibiotic that is effective against most bacteria, filamentous fungi, yeast, plant, and animal cells.
Can zeocin be used as a drug resistance marker?
Since Zeocin™ is active in both bacteria and mammalian cell lines, vectors can be designed that carry only one drug resistance marker for selection. Description. Zeocin™ is a formulation of phleomycin D1, a basic, watersoluble, copper-chelated glycopeptide isolated from Streptomyces verticillus.