Addgene: Protocol - How to Inoculate a Bacterial Culture (2024)

FAQs

Addgene: Protocol - How to Inoculate a Bacterial Culture? ›

Using a sterile pipette tip or toothpick, select a single colony from your LB

Inoculate the liquid with a single bacterial colony by touching a sterile inoculating loop, needle, toothpick, or applicator stick to the colony, making certain that some of the cells have been transferred, and then dipping the inoculating loop, needle, or tip of the stick or toothpick into the liquid and shaking it a ...

Bacteria and other microbes can be inoculated into various media where they thrive. This process of inoculation is carried out in different media such as – plate culture, slant culture, agar plates, stab culture, sweep plate method, and so on.

Bacterial vegetative cells and spores are suspended in sterile tap water or sterile saline, which is then added to the broth. In case of nonsporulating fungi and actinomycetes, hyphae are used for inoculum, and then transferred to the final solution.

Growth of standard expression cultures (100 ml)

Grow the cultures overnight at 37°C. Inoculate 100 ml prewarmed medium (with antibiotics) with 5 ml of the overnight culture and grow at 37°C with vigorous shaking until an OD600 of 0.6 is reached (30–60 min).

If a bacterial culture is left in the same media for too long, the cells use up the available nutrients, excrete toxic metabolites, and eventually the entire population will die. Thus bacterial cultures must be periodically transferred, or subcultured, to new media to keep the bacterial population growing.

At each step, the inoculum is used at 0.5–5% of the medium volume, which allows a 200-fold increase in inoculum volume at every step. Mostly, the inoculum used for the production stage is approximately 5% of the medium volume, or it can be adjusted to obtain maximum fermentor productivity.

What is the direct method of inoculation? ›

In direct inoculation, a small volume of sample is removed aseptically from the sample unit and inoculated directly into a suitable volume of growth medium before incubation. While simple, this method of testing can have some significant limitations.

inoculation, process of producing immunity that consists of introduction of the infectious agent into the body. Historically, inoculation involved introducing the infectious agent onto an abraded or absorptive skin surface.

Using a sterile pipette tip or toothpick, select a single colony from your LB agar plate. Drop the tip or toothpick into the liquid LB + antibiotic and swirl. Loosely cover the culture with sterile aluminum foil or a cap that is not air tight. Incubate bacterial culture at 37°C for 12-18 hr in a shaking incubator.

There are two methods for inoculating (putting the bacteria on) agar plates: quadrant streaking and spread plating. Which one you choose depends on what you want to do after. The quadrant streaking technique is used to isolate single colonies of one or more types of bacteria or fungi.

Some bacteria can be mixed in or sprinkled on the soil during planting. Others need to be revitalized from a dormant state and cultivated by hand—usually by adding a measurement of granules into water.

Inoculation is carried out under aseptic conditions. In this process explants or micro shoots are transferred on to the sterilized nutrient medium. species and sometimes during the various stages of developments. The cultures are observed daily for growth and any signs of infection/ contamination.