Team:TU Munich/Lab
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==In vitro Measurements== | ==In vitro Measurements== | ||
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- | A second possibility to measure parameters of our switches we came up with, was the idea to investigate our system on the transcriptional level only. Therefore, we decided to use malachite green as reporter. Malachite green in a fluorescent dye, whose emission increasing dramaticly (about 3000 times) upon binding of a specific RNA-aptamer.<br><br> | + | A second possibility to measure parameters of our switches we came up with, was the idea to investigate our system on the transcriptional level only. Therefore, we decided to use malachite green as reporter. Malachite green in a fluorescent dye, whose emission increasing dramaticly (about 3000 times) upon binding of a specific RNA-aptamer. |
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[[Image:TUM2010_Malachitgruen-2.png|500px|center|our idea]] | [[Image:TUM2010_Malachitgruen-2.png|500px|center|our idea]] | ||
- | ---concept to be | + | ---concept to be described, as well as literature--- |
<ref>refs</ref> | <ref>refs</ref> | ||
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==In vitro Translation== | ==In vitro Translation== |
Revision as of 19:37, 10 October 2010
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Experiment DesignIn vivo Measurements
Our initial idea to prove our concept of antitermination was to use flourescent proteins as reporters. This approach gives the opportunity to measure the termination and antitermination efficiency of our designed BioBricks in vivo as well as in vitro, the latter using a translation kit based on E. coli lysate.
Wiith these measurements, it should also be possible to observe differences in efficiency of termination as well as antitermination between our designed switches. In vitro Measurements
A second possibility to measure parameters of our switches we came up with, was the idea to investigate our system on the transcriptional level only. Therefore, we decided to use malachite green as reporter. Malachite green in a fluorescent dye, whose emission increasing dramaticly (about 3000 times) upon binding of a specific RNA-aptamer.
---concept to be described, as well as literature---
<ref>refs</ref>
Also we made constructs, where the transcription of the signal-RNA is under the control of a sigma(70) promoter. These two linear DNA-constructs, together with the e.coli RNA-polymerase and the right buffer conditions should represent an easy-to-handle measurement kit on the transcriptional level. Close In vitro Translation
So haben wir in vivo translation gemessen Close
ProtocolsMolecular BiologyPCR
So geht ne PCR CloseDigestion
So geht ne PCR CloseIn vivo measurement
So haben wir in vivo gemessen CloseIn vitro measurement
So haben wir in vitro gemessen Close
Lab BookExplanation:
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