How much G418 to use?

G418 is normally used at a concentration of 400 μg/ml. However, the optimal concentration needs to be determined for your cells.

How long does it take for G418 to kill cells?

3. Replace the G418-containing medium every 2 – 3 days and examine cells for visual toxicity. Most non-transfected (non-resistant) cells will die within 10 days, leaving the transfected cells to expand.

How to prepare G418?

200 mg/ml G-418 Stock Solution:

1. Weight 2gr of G418 (CAS: 108321-42-2 mw=692.71).
2. Add 10ml of sterile H2O.
3. Prewet a 0.22um syringe filter by drawing through 5-10ml pf sterile H2O and discard water.
4. Sterilize G418 Stock through through the prepared 0.22um syringe filter.
5. Stock may be kept at -20C for 1 year.

How is G418 used as a selective agent?

Selection Antibiotics. Commonly know as G418 or G-418, Gibco Geneticin reagent is an aminoglycoside related to gentamicin, and is commonly used as a selective agent for eukaryotic cells. As an analog of neomycin sulfate, it interferes with the function of 80S ribosomes and protein synthesis in eukaryotic cells.

What is Thermo Fisher Scientific G418 reagent used for?

G418, Geneticin | Thermo Fisher Scientific – US G418, Geneticin Commonly know as G418 or G-418, Gibco Geneticin reagent is an aminoglycoside related to gentamicin, and is commonly used as a selective agent for eukaryotic cells.

How much g-418 for selection and maintenance?

The general guideline is 400 µg/mL for selection and 200 µg/mL for maintenance are required for mammalian cells. Plant cells require 25-50 µg/mL for selection and 10 µg/mL for maintenance and bacteria 8-16 µg/mL for selection. Using an analytical balance, weigh an appropriate amount of G-418 powder (use formula below).

How is G418 selection used in cell isolation?

The neomycin (neo) resistance selection procedure (G418 selection) allows for isolation of stable transfectants and is introduced into cells either linked in cis to genes of interest as one DNA fragment, or it can be introduced in trans by co-transfection.