Difference between revisions of "Team:ASTWS-China/InterLab"

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               <h2 class="header">COMMON PAGE</h2>
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               <h2 class="header">InterLab</h2>
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                 Our idea of project comes from Hybridoma technology. We found some disadvantage about the old technology. In order to improve this technology, we started this project. We also want to know how most of people thinks about Hybridoma technology and our new approach. So we make a questioner to collect others view about our project.
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                 Aiming to help develop a reliable and detailed protocol to enable direct florescence data comparison between labs, we decided to collaborate with The Measurement Committee as a Interlab study participant.
 
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                The Interlab study required us to follow a standard protocol including transformation, incubation, plate reader calibration, florescence standard curve obtainment, cell culture florescence measurement, etc ( Detailed protocol information can be found at https://2017.igem.org/Competition/InterLab_Study ). During the whole Interlab study process, everyone, instructed by the team PI, studied the protocol carefully and worked on providing accurate and reliable experiment data to The Measurement Committee.
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              <p>1.Transformation of the test devices and +/- control</p>
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                Unfortunately, we had an unexplained problem while attempting to transform cells with the extracted DNA from the Distribution Kit Plates. Luckily, we heard from iGEM team Worldshaper-XSHS that they have successfully obtained the transformed colony but was unable to continue with the protocol due to lack of a plate reader in their lab. We invited representatives from Worldshaper-XSHS to join our Interlab project, providing transformed colony and they accepted our request. (To learn more about our cooperation, check out our Collaboration[该词链接至collaboration] page.)
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              <p>2.OD600 reference point measurement</p>
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                We strictly followed the standard protocols provided by the Measurement Committee and turned off pathlength correction accordingly.
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              <p>3.Fluorescein standard curve obtainment</p>
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                We measured the fluorescence of 4 replicates of dilution series of fluorescein in PBS buffer. The results are as shown below.
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              <p>4.Cell measurement</p>
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                We had correctly diluted our overnight transformed cell culture to a target OD600 of 0.02. Then we incubated the diluted culture and took samples at 0,2,4,6 hours after dilution and held them on ice. Then we measured the florescence of these samples and the results are as shown below.
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              <p>Raw plate readings</p>
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              <p>Adjusted results</p>
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                To know the opinion from normal people, we started a questionnaire on WeChat.
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Revision as of 08:22, 29 October 2017

InterLab

Aiming to help develop a reliable and detailed protocol to enable direct florescence data comparison between labs, we decided to collaborate with The Measurement Committee as a Interlab study participant.

The Interlab study required us to follow a standard protocol including transformation, incubation, plate reader calibration, florescence standard curve obtainment, cell culture florescence measurement, etc ( Detailed protocol information can be found at https://2017.igem.org/Competition/InterLab_Study ). During the whole Interlab study process, everyone, instructed by the team PI, studied the protocol carefully and worked on providing accurate and reliable experiment data to The Measurement Committee.

1.Transformation of the test devices and +/- control

Unfortunately, we had an unexplained problem while attempting to transform cells with the extracted DNA from the Distribution Kit Plates. Luckily, we heard from iGEM team Worldshaper-XSHS that they have successfully obtained the transformed colony but was unable to continue with the protocol due to lack of a plate reader in their lab. We invited representatives from Worldshaper-XSHS to join our Interlab project, providing transformed colony and they accepted our request. (To learn more about our cooperation, check out our Collaboration[该词链接至collaboration] page.)

2.OD600 reference point measurement

We strictly followed the standard protocols provided by the Measurement Committee and turned off pathlength correction accordingly.

3.Fluorescein standard curve obtainment

We measured the fluorescence of 4 replicates of dilution series of fluorescein in PBS buffer. The results are as shown below.

4.Cell measurement

We had correctly diluted our overnight transformed cell culture to a target OD600 of 0.02. Then we incubated the diluted culture and took samples at 0,2,4,6 hours after dilution and held them on ice. Then we measured the florescence of these samples and the results are as shown below.

Raw plate readings

Adjusted results

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