Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.13 Determining E coli concentration with spectrophotometer

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: '''Small Phage May - June Notebook: Experiments'''</font>

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: [[Team:BYU_Provo/Small_Phage|Overview]]

: [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]]

: [[Team:BYU Provo/Notebook/SmallPhage/Springexp|May-June]]

: [[Team:BYU Provo/Notebook/SmallPhage/Summerexp|July-August]]

: [[Team:BYU Provo/Notebook/SmallPhage/Fallexp|September-October]]

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<font size="5"> '''5.13 Determining E coli Concentration With Spectrophotometer''' </font>

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'''I) Purpose'''
: To determine if OD600 can be used to determine E coli concentration
: According to reference The number of cells in a bacterial culture can be estimated by reading the absorbance at 600nm (OD600). For E. coli cells, an OD600 of 1.0 ≈ 5E8 cells/ml

'''II) Expected Outcome'''
: linear relationship between absorption and 1/dilution factor

'''III) Reagants Used'''
: E. Coli BL21 overnight from 5.12; LB prepared by TA in 100mL flask


'''IV) Actual Procedure'''

1. Dilution series
: Label 6 tubes 1 through 6, to each of the test tubes add 2mL of LB.
: 5mL of E coli overnight (21 hours) was to test tube 1. The content was then pipetted up and down to mix well.
: Performing dilution series by transforming 5mL from tube n to tube n+1 in order of n=1,2,3,4,5. The dilution factor is thus 7/5

2. Measurements using spectrophotometer
: Spectrophotometer was warmed up and set to take measurements at 600nm.
: Pure LB was used to as blank
: Before each measurement, the cuvette was rinsed with 1mL of distilled water and 1mL of E coli solution to be measured. 
: 1mL of E coli solution at various concentration was pipetted into each cuvette for measurement.


'''V) Results'''

1. Raw data

{| class="wikitable"
|+Data From Spectrophotometer Reading
|-
|Test Tube
|Concentration with Respect to Overnight
|Absorption at 600nm
|-
|0
|1
|0.623
|-
|1
|0.714
|0.451
 |-
|2
|0.510
|0.313
|-
|3
|0.364
|0.288
|-
|4
|0.260
|0.182
|-
|5
|0.186
|0.098
|-
|6
|0.133
|0.137
|}

2. Plot

[[File:Spec.png]]

3. Linear Regression Result

ADD

'''VI) Conclusion'''


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