Difference between revisions of "Team:Aalto-Helsinki/InterLab"
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<h3>Cultivations and measurement</h3> | <h3>Cultivations and measurement</h3> | ||
| − | <p>Followed the next protocol for preparing the samples:</br></br>Streaked out LB-plates with E.coli TOP10 strain containing every device and control with chloramphenicol concentration of 35ug/ml. Incubated plates overnight (18-20 hours) at 37C. Picked up biological triplates from the plates and inoculated 3 ml liquid cultures from the colonies with 12ml polypropylene test tube. Incubated the tubes at 37C with shaking at 300rpm for | + | <p>Followed the next protocol for preparing the samples:</br></br>Streaked out LB-plates with E.coli TOP10 strain containing every device and control with chloramphenicol concentration of 35ug/ml. Incubated plates overnight (18-20 hours) at 37C. Picked up biological triplates from the plates and inoculated 3 ml liquid cultures from the colonies with 12ml polypropylene test tube. Incubated the tubes at 37C with shaking at 300rpm for 18 hours. Measured OD600 values of cultures in cuvettes and calculated the dilution required for each sample to obtain the OD value of 0,5. Diluted and re-measured the samples to reach 5% accuracy of 0,5. Transfered 200ul of each sample into 96-well transparent plate and set the platereader to read fluorescence intensity. Proceed with the measurements with the following equipment: </br> </br>Type: Cell Imaging Multi-Mode Plate Reader </br> |
| + | Name and model: Cytation 3 </br> | ||
| + | Company: BioTek Instruments, Inc</br></br> | ||
| + | The following parametres were used:</br></br> | ||
| + | Excitation wavelength: 470 nm</br>Emission wavelength: 511 nm</br>Optics Position: Bottom </br> Filter: Quadruple Monochromators with Bandwith of 16 nm</br> Units Reported: RFU</br></br></p> | ||
<h2>Results</h2> | <h2>Results</h2> | ||
| + | <p>Most samples generated overflow values which could not be measured with the plate reader. The incubation may have generated too much fluorescence proteins even though the samples were diluted properly. The problem wasn't solved after new dilutions of 9:10 ratio and due to the time limit, other ratios weren't attempted.</p> | ||
<h4>1st Technical Replicon </h4> | <h4>1st Technical Replicon </h4> | ||
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</table> | </table> | ||
| + | <h2>Restriction Map</h2> | ||
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Revision as of 12:04, 3 September 2015
