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Team:Warsaw/RBSmeasurement
From 2011.igem.org
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| + | <div class="note">Sample preparation</div> | ||
| + | <div>To measure fluorescence of samples 3 milliliters of LB were inoculated with a colony from the plate. Cultures were vigorously shaken in 37° C overnight. The next step was to centrifuge 1,5 milliliters of liquid culture in an eppendorf tube. Pellet was then resuspended in 100 microliters of RF. Finally, prepared samples were applied on the | ||
| + | fluorimeter plates. We used Corning 96 deep well plates with clear UV transparent bottom. Measurments were carried out using Tecan HS2000 pro fluorimeter with bottom read mode and using active gain optimization | ||
| + | function. Wavlength values for individual proteins: | ||
| + | <dl> | ||
| + | <dt>GFP</dt> | ||
| + | <dd>Excitation 488 nm</dd> | ||
| + | <dd>Emission 509 nm</dd> | ||
| + | <dt>YFP</dt> | ||
| + | <dd>Excitation 514 nm</dd> | ||
| + | <dd>Emission 530 nm</dd> | ||
| + | RFP | ||
| + | Excitation 554 nm | ||
| + | Emission 585 nm | ||
| + | OFP | ||
| + | Excitation 545 nm | ||
| + | Emission 568 nm | ||
| + | SF-GFP | ||
| + | Excitation 488 nm | ||
| + | Emission 508 nm | ||
| + | </div> | ||
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Revision as of 20:46, 17 September 2011
Project description
RBS Measurement
In order to fine-tune expression of genes used in our project we have conducted measurement of various ribosome binding sites included in 2010 spring distribution. Our list of measured parts includes RBSs both from Community and Anderson's collections.
We have used standard measurement kit composed of promoter BBa_J23100 and GFP+terminator part BBa_I130401. All measurements were carried out using flow cytometer.
Results are here
We have used standard measurement kit composed of promoter BBa_J23100 and GFP+terminator part BBa_I130401. All measurements were carried out using flow cytometer.
Results are here
Sample preparation
To measure fluorescence of samples 3 milliliters of LB were inoculated with a colony from the plate. Cultures were vigorously shaken in 37° C overnight. The next step was to centrifuge 1,5 milliliters of liquid culture in an eppendorf tube. Pellet was then resuspended in 100 microliters of RF. Finally, prepared samples were applied on the
fluorimeter plates. We used Corning 96 deep well plates with clear UV transparent bottom. Measurments were carried out using Tecan HS2000 pro fluorimeter with bottom read mode and using active gain optimization
function. Wavlength values for individual proteins:
- GFP
- Excitation 488 nm
- Emission 509 nm
- YFP
- Excitation 514 nm
- Emission 530 nm RFP Excitation 554 nm Emission 585 nm OFP Excitation 545 nm Emission 568 nm SF-GFP Excitation 488 nm Emission 508 nm
