What does hygromycin B do?
Hygromycin B is a water-soluble antibiotic purified from the bacterium Streptomyces hydroscopicus. Hygromycin B acts by binding to the 80S subunit of the bacterial ribosome and inhibiting protein synthesis. Gibco® Hygromycin B is used as a bacterial selection antibiotic in the concentration range of 200–500 µg/mL.
How long does Hygromycin last?
The antibiotic is stable for at least 2 years at 4°C. It is stable for about one month at 37°C.
How long do Hygromycin plates last?
ANSWER: Hygromycin B solution is stable for at least two years at +4°C and about one month at +37°C.
How long does it take for hygromycin selection?
Ideally, control cells should die within one week after addition of the antibiotic, allowing colonies of resistant cells to form within 10–14 days. Hygromycin B is added to the culture medium at a concentration that varies with the cell type transfected.
How does hygromycin selection work?
Hygromycin is active against both prokaryotic and eukaryotic cells. It acts by inhibiting polypeptide synthesis. It stabilizes the tRNA-ribosomal acceptor site, thereby inhibiting translation.
How do you dilute hygromycin?
Stock Solution and Working Concentrations o Use this protocol to prepare either a stock solution of 50 or 100 mg/ml Hygromycin. o Dilute to the following concentrations, based on the application: • For bacteria selection: 20-200 µg/ml. For plant cell maintenance: 20 µg/ml. For plant cell selection: 20-200 µg/ml.
How do you dissolve hygromycin B?
Dissolve Hygromycin B in 1 mL of distilled water. NOTE: If not used immediately, solution may be stored at 2 – 8°C for at least 4 weeks. Alternatively, aliquot and store at -20°C. After thawing the aliquots, do not re-freeze.
Is hygromycin B light sensitive?
The molecules of hygromycin are still light sensitive, no matter how much is there… you need to protect it from light.
How do you make a hygromycin stock solution?
Hygromycin (40 mL) For a stock solution of 5 mg/mL: 200 mg Hygromycin. 40 mL ddH2O. Filter sterilize.
Is hygromycin light sensitive?
How do you store hygromycin B?
Since solutions are stable refrigerated, freezing should be avoided. Hygromycin B products should be stored as supplied at 2-8° C. The dry solid is stable for at least five years if stored at 2-8 °C.
Can hygromycin B be autoclaved?
Hygromycin B is soluble in water, at concentrations >50 mg/mL, and stable in aqueous solution for at least five years at 2-8°C. methanol or ethanol. found to remain stable for at least a month at 37°C and Page 2 60°C. Solutions should be sterilized by filtration, not autoclaving.
Does hygromycin B kill non transfected cells?
Hygromycin B is added to the culture medium at a concentration that varies with the cell type transfected. A titration experiment for each cell type may therefore be performed to determine the amount of Hygromycin B needed to kill non-transfected cells.
Is hygromycin B prokaryotic or eukaryotic?
Cloning of the resistance gene and fusion with eukaryotic promoters has resulted in the development of vectors that permit selection for resistance to Hygromycin B in both prokaryotic and eukaryotic cells. What is the stability of Hygromycin B in solution? The antibiotic is stable for at least 2 years at 4°C.
Why do we need to replace media with hygromycin B?
To maintain Hygromycin-resistant phenotype of transfected cell lines and for the elimination of revertants, cells may be regularly cultured in medium containing Hygromycin B at the same concentration used for the initial selection. When is replacement of media required?
What is the mechanism of resistance to hygromycin?
The resistance gene codes for a kinase that inactivates the antibiotic through phosphorylation. Cloning of the resistance gene and fusion with eukaryotic promoters has resulted in the development of vectors that permit selection for resistance to Hygromycin B in both prokaryotic and eukaryotic cells.