Difference between revisions of "Team:CMUQ/Biofilm Formation Lab R"

 
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; "><b> 2. Formation of Biofilm of SRB8 and DH5 at optimum NaCl concentration range (1.7-2.5 M) in 2X LB </b></p>
 
<p style="font-size: 20px;margin-right:40px; margin-left:40px; "><b> 2. Formation of Biofilm of SRB8 and DH5 at optimum NaCl concentration range (1.7-2.5 M) in 2X LB </b></p>
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<img src="https://static.igem.org/mediawiki/2017/5/59/Biofilmr6.png" style="margin-left:140px;" width=55%>
 
<img src="https://static.igem.org/mediawiki/2017/5/59/Biofilmr6.png" style="margin-left:140px;" width=55%>
 
<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">SRB8 cells were treated with 2X LB medium with different NaCl concentrations and plated on a tissue culture treated 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation. </p>
 
<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">SRB8 cells were treated with 2X LB medium with different NaCl concentrations and plated on a tissue culture treated 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation. </p>
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<p style="font-size: 24px;margin-right:40px; margin-left:40px;color:#F28C19; "><b> Analysis:</b></p>
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<p style="font-size: 20px;margin-right:40px; margin-left:60px; color:#F28C19; ">Figure 1: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">An increase in the absorbance was observed from 0 M NaCl to 0.2 M NaCl. The maximum absorbance for DH5 in artificial sea water was at 0.2 M NaCl. Then the absorbance decreased from 0.2 M NaCl to 0.8 M NaCl. Followed by an increase through 0.8 M NaCl to 1.6 M NaCl. </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:60px; color:#F28C19; ">Figure 2: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">An increase in the absorbance was observed from 0 M NaCl to 0.1 M NaCl. The maximum absorbance for DH5 in 2X LB was at 0.1 M NaCl. After 0.1 M NaCl the absorbance started fluctuating through NaCl concentrations 0.1 M to 1.6 M. </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:60px; color:#F28C19; ">Figure 3: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">An increase in the absorbance was observed from 0.05 M NaCl to 0.2 M NaCl. The maximum absorbance for SRB in artificial sea water was at 0.2 M NaCl. After 0.2 M NaCl the absorbance started fluctuating through NaCl concentrations 0.2 M to 1.6 M.</p>
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<p style="font-size: 20px;margin-right:40px; margin-left:60px; color:#F28C19; ">Figure 4: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">A decrease in the absorbance was observed from 0 M NaCl to 0.4 M NaCl. The maximum absorbance for SRB in 2X LB was at 0.8 M NaCl. Then the absorbance decreased from 0.8 M NaCl to 1.6 M NaCl. </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:60px; color:#F28C19; ">Figure 5: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">The absorbance was fluctuating through the various NaCl concentrations within the range of 1.7 to 2.5 M NaCl. The maximum absorbance for DH5 in 2X LB was at 0 M NaCl</p>
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<p style="font-size: 20px;margin-right:40px; margin-left:60px; color:#F28C19; ">Figure 6: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; ">The absorbance was fluctuating through the various NaCl concentrations within the range of 1.7 to 2.5 M NaCl. The maximum absorbance for SRB in 2X LB was at 2.1 M NaCl </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; > Based on the results obtained, there is no direct relationship between NaCl concentration and biofilm formation.</p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px; color:#F28C19; ">Anomalies: </p>
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<p style="font-size: 20px;margin-right:40px; margin-left:40px;>High values of standard deviation indicate experimental errors associated with preparing and plating the cultures. Difference in replicates’ absorbance is due to unspecific binding of the crystal violet the walls of the wells of 96 well plates. Also, errors in pipetting might have contributed to different absorbance in replicates or contamination of the plate itself. </p>
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Latest revision as of 00:47, 2 November 2017

Biofilm Lab Results

Biofilm formation was tested under conditions similar to that of seawater conditions using artificial seawater prepared in lab, which contains main salts and concentrations found in seawater. This test was done as a control to observe biofilm formation under standard conditions of artificial seawater.

Results indicated that biofilm was formed since absorbance is directly related to biofilm formation, and any increase in the absorbance is directly related to biofilm formation. We have found also that DH5 is capable of forming biofilm.

Different samples Average Absorbance Standard Deviation
DH5 0.2615 0.07797649
SRB5A 1.1185 0.16140529
SRB7 1.3985 0.13931619
SRB8 1.44375 0.25075137

1. Minimal formation of Biofilm of SRB8 and DH5 at lower NaCl concentration range (0.05-1.6 M) in artificial sea water and 2X LB

SRB8 DH5
NaCl Concentration (M) Average Absorbance in Sea water Average Absorbance in 2X LB Average Absorbance in Sea water Average Absorbance in 2X LB
0 0.767666667 1.462666667 0.950333333 3.703333333
0.5 1.739 1.418 1.035333333 4.210666667
0.1 1.1 0.936 1.585666667 4.278333333
0.2 1.153666667 0.850333333 1.793666667 4.133666667
0.4 1.075333333 0.717666667 1.187333333 2.028666667
0.8 1.042333333 1.658666667 1.142666667 2.156666667
1.6 0.843666667 0.658666667 1.926333333 1.350333333

DH5 cells were treated with artificial sea water with different NaCl concentrations and plated on a 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation.

DH5 cells were treated with 2X LB medium with different NaCl concentrations and plated on a 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation.

SRB8 cells were treated with artificial sea water with different NaCl concentrations and plated on a 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation.

SRB8 cells were treated with 2X LB medium with different NaCl concentrations and plated on a 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation.

2. Formation of Biofilm of SRB8 and DH5 at optimum NaCl concentration range (1.7-2.5 M) in 2X LB

NaCl Concentration (M) SRB8 DH5
0 0.129666667 0.194666667
1.7 0.099 0.088
1.9 0.143666667 0.092333333
2.1 0.150333333 0.111333333
2.3 0.090666667 0.077333333
2.5 0.088333333 0.076333333

DH5 cells were treated with 2X LB medium with different NaCl concentrations and plated on a tissue culture treated 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation.

SRB8 cells were treated with 2X LB medium with different NaCl concentrations and plated on a tissue culture treated 96-well plate. The plate was incubated at 37C for ~48 hours. This was followed by staining with 0.1% crystal violet solution to measure the absorbance which corresponds to the biofilm formation.

Analysis:

Figure 1:

An increase in the absorbance was observed from 0 M NaCl to 0.2 M NaCl. The maximum absorbance for DH5 in artificial sea water was at 0.2 M NaCl. Then the absorbance decreased from 0.2 M NaCl to 0.8 M NaCl. Followed by an increase through 0.8 M NaCl to 1.6 M NaCl.

Figure 2:

An increase in the absorbance was observed from 0 M NaCl to 0.1 M NaCl. The maximum absorbance for DH5 in 2X LB was at 0.1 M NaCl. After 0.1 M NaCl the absorbance started fluctuating through NaCl concentrations 0.1 M to 1.6 M.

Figure 3:

An increase in the absorbance was observed from 0.05 M NaCl to 0.2 M NaCl. The maximum absorbance for SRB in artificial sea water was at 0.2 M NaCl. After 0.2 M NaCl the absorbance started fluctuating through NaCl concentrations 0.2 M to 1.6 M.

Figure 4:

A decrease in the absorbance was observed from 0 M NaCl to 0.4 M NaCl. The maximum absorbance for SRB in 2X LB was at 0.8 M NaCl. Then the absorbance decreased from 0.8 M NaCl to 1.6 M NaCl.

Figure 5:

The absorbance was fluctuating through the various NaCl concentrations within the range of 1.7 to 2.5 M NaCl. The maximum absorbance for DH5 in 2X LB was at 0 M NaCl

Figure 6:

The absorbance was fluctuating through the various NaCl concentrations within the range of 1.7 to 2.5 M NaCl. The maximum absorbance for SRB in 2X LB was at 2.1 M NaCl

Anomalies: