Team:Warsaw/Calendar-Stage1/29 August 2010

From 2010.igem.org

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<br><br>Note:We improved the way of cell preparation by using lysozyme or triton to reduce the adherence of cells
<br><br>Note:We improved the way of cell preparation by using lysozyme or triton to reduce the adherence of cells
<br><br><div class="note"> Previous 2 days: Set up and culturing Hela cell culture [Kamil]</div>
<br><br><div class="note"> Previous 2 days: Set up and culturing Hela cell culture [Kamil]</div>
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<br> Use a protocol...
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<br> Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (<a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_59:_Quantitative_measurement_of_mamallian_cell_invasion_by_bacteria_using_flow_cytometry">RFC 59 standard</a> point 5)
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<br>
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<br><div class="note"> Previous day: Set up liquid cultures of different types of E.coli [Marta B.]</div>
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<br><div class="note"> Previous day: Set up liquid cultures of different types of E.coli [Ania P.]</div>
-
<br> Use a protocol...
+
<br> Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (<a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_59:_Quantitative_measurement_of_mamallian_cell_invasion_by_bacteria_using_flow_cytometry">RFC 59 standard</a> point 5)
<br>
<br>
<br><div class="note"> Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]</div>
<br><div class="note"> Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]</div>
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<br>1. Incubation of Hela cells with different E.coli types
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<br>1. Incubation of Hela cells with different E.coli types.
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<br>2. Preparation of probes with incubated cells (using lysozyme and Triton) and examining them in flow cytometer
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<br>2. Preparation of probes with incubated cells and examining them in flow cytometer.
-
<br>
+
<br>Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (<a href="http://openwetware.org/wiki/The_BioBricks_Foundation:RFC#BBF_RFC_59:_Quantitative_measurement_of_mamallian_cell_invasion_by_bacteria_using_flow_cytometry">RFC 59 standard</a> points 6 and 7)<br>
<br><div class="note"> Data analysis with Cell Quest and Microsoft Excel [Cherry]</div>
<br><div class="note"> Data analysis with Cell Quest and Microsoft Excel [Cherry]</div>
<br>
<br>

Revision as of 18:27, 27 October 2010

Example Tabs


30.08.2010 Monday

Note:We improved the way of cell preparation by using lysozyme or triton to reduce the adherence of cells

Previous 2 days: Set up and culturing Hela cell culture [Kamil]

Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (RFC 59 standard point 5)

Previous day: Set up liquid cultures of different types of E.coli [Ania P.]

Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (RFC 59 standard point 5)

Testing the invasivnes of E.coli in Hela cells [Kamil,Cherry]

1. Incubation of Hela cells with different E.coli types.
2. Preparation of probes with incubated cells and examining them in flow cytometer.
Conducted according to the protocol included in "Quantitative measurement of mamallian cell invasion by bacteria using flow cytometry" standard (RFC 59 standard points 6 and 7)

Data analysis with Cell Quest and Microsoft Excel [Cherry]


legend:

E.coli constructs:
G- GFP
J- JPH+Listeriolysin O+Invasin+GFP
A- APH+Listeriolysin O+Invasin+GFP
TOP- without construct

LIZ- treated with lysozyme
TRIT- treated with triton
X- untreated

bacteria:
30-08-2010.001- TOP
30-08-2010.003- G
30-08-2010.004- J
30-08-2010.005- A

Hela cells with bacteria:
30-08-2010.033- X TRIT
30-08-2010.034- A TRIT
30-08-2010.035- J TRIT
30-08-2010.036- A LIZ
30-08-2010.037- J LIZ
30-08-2010.038- without bacteria TRIT
30-08-2010.039- TOP TRIT
30-08-2010.040- G TRIT
30-08-2010.041- X LIZ
30-08-2010.042- without bacteria LIZ
30-08-2010.043- TOP LIZ
30-08-2010.044- G LIZ



Cytometer data from Cell Quest analysis:


Lysozyme treatment:


Triton treatment:





Data from Cell Quest processed with Microsoft Excel:



TOP- Hela cels with regular TOP E.coli
M1,M2,M3,M4- %of events gated with markers M from total