Team:UNIPV-Pavia/Methods Materials/Absorbance

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(→Different protocols for different growth curves)
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As our first step we wanted to individuate a proper protocol to have reproducible growth curves for the coltures incubated inside the microplate reader.  
As our first step we wanted to individuate a proper protocol to have reproducible growth curves for the coltures incubated inside the microplate reader.  
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We compared three different solution, as you can see in the figures below.
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''Protocol 1'':
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*overnight colture incubation in 5ml LB+Amp 37°C 220 rpm; 
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*dilution 1:1000;
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*incubation 37°C 220 rpm for about 4 hours.
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''Protocol 2'':
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''Protocol 3'':
[[Image:unipv_asb_protocols_12.jpg| 500px|Growth Protocols n.1 and n.2|thumb|center|Growth Protocols n.1 and n.2]]
[[Image:unipv_asb_protocols_12.jpg| 500px|Growth Protocols n.1 and n.2|thumb|center|Growth Protocols n.1 and n.2]]

Revision as of 10:42, 17 October 2009

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Measurements - Absorbance


Absorbance and water dispensation

We find no relevant differences between the measures before and after the dispensation of different volumes of water in the well, both for bacterial coltures and simple liquid growth medium.

You can find more informations about the experiment at [http://aimed11.unipv.it/iGEM2009 Download Protocol] - Test n.9bis, 10/10/09.

You can find more informations about the experiment at [http://aimed11.unipv.it/iGEM2009 Download Protocol] - Test 17/10/09.


Absorbance and water dispensation

Absorbance and volume

The proportionality between absorbance (OD600) measurement and volume of colture in the well was verified. By fitting a linear regression model on the experimental data we estimate a coefficient of proportionality of 8.27e-4.

You can find more informations about the experiment at [http://aimed11.unipv.it/iGEM2009 Download Protocol] - Test n.9bis, 10/10/09.


Absorbance and volume

Absorbance and dilutions in liquid growth medium

The proportionality between absorbance measurement (OD600) and colture's dilutions was verified, confirming the possibility of using OD600 to measure the bacterial quantity in the well, regardless of the total volume present in the well.

This experiment was done in two different growth mediums, LB and M9, and in three different total volumes in the well, 100μl, 200μl and 300μl.

By fitting a linear regression model on the experimental data we estimate a coefficient of proportionality of 0.0018 for LB medium and 0.0035 of for M9.

You can find more informations about the experiments at [http://aimed11.unipv.it/iGEM2009 Download Protocol] - Test n.20, 13/08/09 and Test n.20 "M9", 28/08/09.


Absorbance and colture's dilutions in LB
Absorbance and colture's dilutions in M9

Different protocols for different growth curves

As our first step we wanted to individuate a proper protocol to have reproducible growth curves for the coltures incubated inside the microplate reader.

We compared three different solution, as you can see in the figures below.

Protocol 1:

  • overnight colture incubation in 5ml LB+Amp 37°C 220 rpm;
  • dilution 1:1000;
  • incubation 37°C 220 rpm for about 4 hours.

Protocol 2:

Protocol 3:

Growth Protocols n.1 and n.2
Growth Protocol n.3

Growth ambients in comparison

Growth ambients in comparison


Growth ambients in comparison, exponential phase
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