This experiment aims to place the virtue of utilizing pour home base method and spread method. The growing of the bacteriums will find the whether the experiment is successful. The cogency scope will be 30-300 settlement organizing unit with merely one type of settlement. The unsuccessful consequence is when there is no colony signifier ; less or more than the cogency scope or more than one type of settlement formed ( cfu ) .

Dilution is critical as the unknown sample may hold more than one cell. By consecutive thining an being in an agar home base can be reached where merely one cell remains in the medium. In the terminal, a pure civilization is obtained. Besides, from an ab initio high concentration, the cell concentration is decreased. Cell concentration in a sample can be in a scope of 1000 and 1000000s and even one million millions. Therefore, it makes good sense to thin the sample. One effectual method without utilizing excessively much dilutant is by consecutive dilution. A procedure of thining a sample is by executing a series of perennial dilutions. The consecutive dilution can be in the signifier of 2-fold, 5-fold, 10-fold or even 1000-fold dilution, depending on the cell concentration of the sample.

10ml valve with 9ml of saline or unfertile H2O. Shake the undiluted active dried barm. Then inoculate 1ml of active dried barm and set in the 9ml of saline or unfertile H2O. Shake the solution and label 10-1. Shake the 10-1 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-2. Shake the 10-2 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-3. Shake the 10-3 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-4. Shake the 10-4 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-5. Shake the 10-5 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-6. Shake the 10-6 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-7. Shake the 10-7 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-8. Shake the 10-8 and inoculate 1ml of active dried barm and put in another the 9ml of saline or unfertile H2O. Shake the solution and label 10-9.

4.2 Plating utilizing Spread Plate

Using 0.1ml of 10-6, 10-7 and 10-8 dilution spread onto extra home bases of malt agar utilizing a “ hockey stick ” . When experiencing frictional force playing, halt the spreading. Incubate the home base for inverted at room temperature ( 25EsC ) for 2 yearss. After incubation, choose merely plate that contain between 30-300 settlements per home base and work out the figure of barm cell as cfu/g

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4.3 Plating utilizing pour home base

Pipette 1ml of 10-7, 10-8 and10-9 dilution into extra Petri dish and add 15ml of liquefied malt agar ( app 45EsC ) and mix gently and let the agar to put. Incubate the home base for inverted at room temperature ( 25EsC ) for 2 yearss. After incubation, choose merely plate that contain between 30-300 settlements per home base and work out the figure of barm cell as cfu/g

Consequence and Calculation

Method

Dilution

Number of colonies/plate*

cfu/g

Home plate 1

Home plate 2

Average

Spread home base

10-6

61

50

55.5

5.55X106

10-7

TNTC

TNTC

TNTC

TNTC

10-8

TNTC

TNTC

TNTC

TNTC

Pour home base

10-7

80

TFTC

Nothing

Nothing

10-8

TFTC

TFTC

TFTC

TFTC

10-9

TFTC

TFTC

TFTC

TFTC

Note *TNTC- excessively legion to number, transcending 250 colonies/plate

*TFTC- excessively few to number, below 25 colonies/plate

**Report cfu/g to 1 denary topographic point.

Cell per ml= cell count X dilution factor Ten volume

Spread plate=55.5 settlements X 106 X ( 1/0.1 )

=5.55X108 barm cells

Discussion

Merely dispersed home base of dilution of 10-6 is in scope. The remainder of the home base of both dispersed home base and pour home base is invalid. One of the jobs is the dilution prepared by me and my spouse. The taint by non altering the pipette tips lead to the complete growing of the barm cells.

6.1 Problems of pour home base method

Restriction of the micro-organism

Aerobic micro-organism may non be suited as the growing is mostly in the agar instead on the surface of the agar.

Longer incubation period

Colonies in the agar may non be seeable until subsequently. The settlements in the agar are in partly anaerobiotic status. Facultative anaerobes will therefore grow easy and therefore, will non be really obvious after a twenty-four hours of incubation.

Under-estimation of the cell Numberss

Some barm settlements possibly overlapping one another and may be seen as a individual CFU. Furthermore the cells are exposed to warm molten agar and this can kill some heat-sensitive barm and give a lower CFU home base count.

Problem of spread home base

Some barm settlements possibly overlapping one another and may be seen as a individual CFU.

Decision

This is a successful experiment although the dilution is cross- contaminated. This is because both the spread and pour home bases use the diluted sample which the spread home base shows a valid settlement formed. In the hereafter when managing with yeast sample, I will plate the barm utilizing dispersed home base method. This is chiefly because that barm is heat- sensitive being and aerophilic fungus.

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