LAB 3 (NOR SHAQIRA BT AZLAN)


LAB 3 : PREPARATION & STERILIZATION OF CULTURE MEDIA


INTRODUCTION

growth medium or culture medium is a liquid or gel designed to support the growth of microorganisms ,cell or small plants.
There are two major types of growth media: those used for cell culture, which use specific cell types derived from plants or animals, and microbiological culture, which are used for growing microorganisms, such as bacteria or yeast. The most common growth media for microorganisms are nutrient broths and agar plates, a specialized media are sometimes required for microorganism and cell culture growth.

figure : agar culture media


Autoclaves are used in sterilization. The advantage of using an autoclave is that it can reach temperatures higher than boiling water alone, so it can kill not only bacteria but also bacterial spores, which tend to be resistant. Autoclaves are used in laboratories to assure items such as glassware and surgical equipment are sterile.
Autoclaves are built around the principal that the boiling point of water increases when it is under pressure. At 15 pounds of pressure per square inch, the boiling point of water increases from 100 degrees Celsius to 121 degrees Celsius. At this temperature, all life forms are killed within 15 minutes.

figure : autoclave machine




OBJECTIVE

To prepare sterile nutrient agar for culturing microorganisms.




DISCUSSION

The culture media are prepared by weighing all the broth component as below ;

0.6 g/L “Lab-lemco” powder (beef extract)
0.4 g/L yeast extract
1.0 g/L peptone (nitrogen source)
1.0 g/L sodium chloride
3.0 g/L agar powder

figure : agar powder and beef extract

figure : yeast extract

figure : peptone

figure : sodium chloride


Then, put the broth into Scott bottles and dissolve it with 200 mL of distilled water.


                Autoclave sterilization times and temperatures are dependent on the composition and nature of the items being autoclaved.  Minimum times are usually 15 minutes at 250 F (121 C) or 5 minutes at 270 F (132.2 C).  Autoclaves should be set at 250F when sterilizing . Items to be sterilized are loaded into the autoclave in a manner so that nothing touches the inside of the chamber. Items to be sterilized should be placed into the autoclave so that steam can uniformly flow between items. When the autoclave cycle is complete and the pressure returns to zero the door may be opened. Before opening the door,   wear  heat-resistant autoclave gloves.  To prevent being scalded from the steam, move to the opposite side of the door opening.  Open door slowly and once opened, walk away from the direct line of the steam.  After a few minutes, the autoclave materials can then be removed.  Items will still be very hot for a while.  Items should not be used until cool to the touch.



The benefit of autoclaving is it  allows materials to be sterilized within a relatively short time frame without the use of reagents.


CONCLUSION

The media culture prepared let microorganisms grow safely in an ensured environment that has all the important nutrients and to protect them against any environmental damage so these organisms can be used when needed.







REFERENCES

en.wikipedia.org/wiki/Growth_medium

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