Bacterial Culture Techniques - Essay Example

BACTERIA CELL CULTURE TECHNIQUE DEFINITION AND EXPLANATION BACTERIA Bacteria are organisms that are single celled and hence cannot be seen with the naked eye, the live in and around all of us and can be helpful but in many other cases they can cause illness people some of the common illnesses cause by Bacteria are ear infections, strep throat and pneumonia hogenic in extreme cases. Bacteria are known to live in water or in all types of dirt also in water and also to live with in humans and animals where they can grow and spread.

Bacteria is said to be found naturally growing in side the human body and humans are known to be able to contract bacteria from other people through conditions such as unprotected sex etc. Bacteria have been often termed as being the primitive beings and are said to help with the nitrogen cycle. In a wider view if looked at bacteria are all those unicellular organisms that belong to the category of Schizomycetes they may have a difference in their requirements for oxygen and nutrients and have difference in morphology as well as have varying motility and be free-living.

Bacteria are also known as prokaryotes in general and these are known to be grouped together as they all do not have nuclear membranes.CULTURE: The growing of cells in a synthetic environment is known as cell culture. That could very well refer to either types of cells be those prokaryotic or eukaryotic cells. Culture can also be called the in vitro growing of cells of either plants or animals in a nutrient artificial medium. In the process of cell culture the cells used are no longer in an organized tissue form rather they are separate and grown in a simulated environment.

The general steps that are used during cell cultures are as follows:1. The mixture should be stirred well and at a temperature of 120C it should be autoclaved.2. After that the mixture should be poured into a plate and allowed to cool.3. On a stick a small amount of dust should be taken.4. The stick containing the dust should be run through the plate containing the mixture.5. The plate with dust and mixture should be covered6. The incubator should be set at 37C 7. The plate covered, containing both mixture and dust should be placed in the incubator.8. That should be closed and left over night.

BACTERIA CELL CULTURING:Bacteria cell culturing requires the following steps and procedures to be taken care of:APPARATUS and EQUIPMENT:The materials that are necessary for culturing bacteria cells are (a) culture tubes that are made of glass and that have their own labels and metal covers. (b) Media room or customized growth medium for simulation. (c) Para Film is also needed and (d) Pipette tubes that are also made of glass. Other necessary equipment includes Bunsen burners, motorized pipettes and micropipettes along with sterile tips.

THE CULTURE PROCEDURE: The first step in culturing bacteria cells is to streak an Agar plate and then incubate that until there begins a growth in colonies. Some bacteria have a temperature sensitive mutation rate and there fore would require the incubators to be set at 30C however in the case of E.coli the desired temperature for incubation is 37C. In order to be certain that the beginning of this culture has been from a single population of cells streaking of the plate is necessary. It is not necessary that you use an Agar plate as they are only the standard form, other types of plates can also be used.

The next step would be to take the antibodies out of the cool storage area and allow them to melt (thaw) out. Then starting with the glass pipette flame that along with the mouth of the medium bottle that you are going to use. Take a measure of the amount that u need and recap the bottle as soon as possible. Then take the culture tube, remove its cap and flame the top and the pipette and then recap it. Taking the pate with the growths, place the tip of the pipette on the top side sucking up at least one of the colonies and then take it to the culture tube mouth.

Let the single colony eject into the tube, flame both the tube mouth and the pipette and then recap the tube. After that is done with another pipette take the desired amount of antibiotic and slowly eject that into each culture tube then using the vortex mix each tube well. Take those tubes then to the incubator and slowly using the knob slow its speed till it reaches a dead halt then place you tubes in a balanced way. Turn the rotation of the incubator to 7 and turn the incubator back on. Even though for miniprep of plasmid DNA the time interval of over night is sufficient yet the amount of time to wait would depend on the reason that you are growing the cells.

(Porter, Roy.) Keeping in mind that this is a contained experiment one which is based on controlled odds and factors it is essential that the substances that are being used and the results be one hundred percent accurate and they should be such results that come from the mixing of only and only the desired substance that are added to the mixture and not the results of any other foreign substance mixing with the germ/bacterial mixture. The only way to ensure accurate results is the one keeps a close check on procedures and contamination issues, the pre heating of the incubator.

The adding of the mixture the streaking and the agar plate being clean and with out any previous substance still remaining on it. The opening and closing of the bottle and the fire of the pipettes all need to be carefully monitored and conducted. Some bacteria take more time to let their cells culture and keeping in mind the exact bacteria being used one should accordingly calculate the necessary time for incubation, the required amounts of external substances and antibiotics as well as the required level of lighting etc.

The above mentioned techniques and the steps given are essential and are to be followed in accordance with the needs of the type of bacteria being used to ensure proper cell culture taking place. SOURCES:Porter, Roy. The Greatest Benefit to Mankind: A Medical History of Humanity. New York: W.W. Norton, 1998.