Team:Freiburg/Project/toolkit
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<p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/1"> Abstract & Intro </a></p> | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/1"> Abstract & Intro </a></p> | ||
+ | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/crrna"> Targeting </a></p> | ||
<p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/effector"> Effectors </a></p> | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/effector"> Effectors </a></p> | ||
<p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/induction"> Effector Control </a> </p> | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/induction"> Effector Control </a> </p> | ||
- | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/ | + | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/modeling"> Modeling </a></p> |
+ | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/truncation"> Truncation </a></p> | ||
<p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/method"> uniBAss </a></p> | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/method"> uniBAss </a></p> | ||
+ | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/unibox"> uniBOX </a></p> | ||
<p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/toolkit" class="active"> Manual </a></p> | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/toolkit" class="active"> Manual </a></p> | ||
- | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/ | + | <p class="first_order"><a href="https://2013.igem.org/Team:Freiburg/Project/application" > Application </a></p> |
</div> | </div> | ||
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</ul> | </ul> | ||
- | By the end of the routine you will get a personal manual. All you need to | + | By the end of the routine you will get a personal manual. All you need to do is using the uniCAS toolkit and follow the instructions. Best of all: The uniCAS toolkit is all open source and in iGEM standard! |
- | + | ||
- | + | ||
<br> | <br> | ||
- | |||
+ | <!-- | ||
+ | <p><b>Finally ... does everybody know what time it is? It's tooltime!</b></p> | ||
Also have a look at Sigma Aldrich to order your oligos coding for the crRNAs: <br> | Also have a look at Sigma Aldrich to order your oligos coding for the crRNAs: <br> | ||
<br> | <br> | ||
<a href="http://www.sigmaaldrich.com/germany.html"> <img src="https://static.igem.org/mediawiki/2013/0/01/SA_Logo_freiburg.jpg" width="300px"> </a> | <a href="http://www.sigmaaldrich.com/germany.html"> <img src="https://static.igem.org/mediawiki/2013/0/01/SA_Logo_freiburg.jpg" width="300px"> </a> | ||
+ | --> | ||
</div> | </div> | ||
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<div id="check-1" class="toHide" style="display:none"> | <div id="check-1" class="toHide" style="display:none"> | ||
<img src="https://static.igem.org/mediawiki/2013/6/6e/Activation.png"> | <img src="https://static.igem.org/mediawiki/2013/6/6e/Activation.png"> | ||
- | <p> Choose an effector to activate your genes. Click <a id="link" href="https://2013.igem.org/Team:Freiburg/Project/effector#activation"> here </a> to see the functional tests of the different activation effectors. </p> | + | <p> Choose an effector to activate your genes. Click <a id="link" href="https://2013.igem.org/Team:Freiburg/Project/effector#activation"> here</a> to see the functional tests of the different activation effectors. </p> |
<label><input id="rdb4" type="radio" name="toggler_two" value="3" onClick="pageScroll()"/>VP16 (recommended) </label> | <label><input id="rdb4" type="radio" name="toggler_two" value="3" onClick="pageScroll()"/>VP16 (recommended) </label> | ||
<!-- <label><input id="rdb5" type="radio" name="toggler_two" value="4" onClick="pageScroll()"/> ?????? </label> --> | <!-- <label><input id="rdb5" type="radio" name="toggler_two" value="4" onClick="pageScroll()"/> ?????? </label> --> | ||
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<div id="check-2" class="toHide" style="display:none"> | <div id="check-2" class="toHide" style="display:none"> | ||
<img src="https://static.igem.org/mediawiki/2013/d/da/Repression.png"> | <img src="https://static.igem.org/mediawiki/2013/d/da/Repression.png"> | ||
- | <p> Effectively repress your genes using KRAB or | + | <p> Effectively repress your genes using KRAB or G9a as functional effector. <br> Click <a id="link" href="https://2013.igem.org/Team:Freiburg/Project/effector#repression"> here</a> to see the functional tests of the different activation effectors. </p> |
<div> | <div> | ||
<label><input id="rdb4" type="radio" name="toggler_two" value="5" onClick="pageScroll()"/>KRAB </label> | <label><input id="rdb4" type="radio" name="toggler_two" value="5" onClick="pageScroll()"/>KRAB </label> | ||
- | <label><input id="rdb5" type="radio" name="toggler_two" value="6" onClick="pageScroll()"/> | + | <label><input id="rdb5" type="radio" name="toggler_two" value="6" onClick="pageScroll()"/> G9a </label> |
</div> | </div> | ||
</div> | </div> | ||
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<p> | <p> | ||
- | You have chosen to activate a gene using a non inducible dCas9-VP16 device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | + | You have chosen to activate a gene using a non inducible dCas9-VP16 device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). |
</p> | </p> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/7/7c/1_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/2/2b/2_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
- | You have chosen to activate a gene using a UVB light inducible dCas9-UVR8 & COP1-VP16 device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | + | You have chosen to activate a gene using a UVB light inducible dCas9-UVR8 & COP1-VP16 device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). |
</p> | </p> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/d/dd/3_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
- | You have chosen to repress a gene using a non inducible dCas9-KRAB device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | + | You have chosen to repress a gene using a non inducible dCas9-KRAB device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). |
</p> | </p> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/d/dc/4_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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You have chosen to repress a gene using a non inducible dCas9-KRAB device. Therefore you have to order the | You have chosen to repress a gene using a non inducible dCas9-KRAB device. Therefore you have to order the | ||
- | following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our | + | following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our |
plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | ||
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All plasmids are optimized for expression in mammalian systems. The devices are also available containing | All plasmids are optimized for expression in mammalian systems. The devices are also available containing | ||
- | a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> | + | a SV40 instead of a CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> |
side). This system was tested mainly in CHO-K1 and HEK-293T cells. | side). This system was tested mainly in CHO-K1 and HEK-293T cells. | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/0/08/5_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
- | You have chosen to repress a gene using a UVB light inducible dCas9-UVR8 & COP1-KRAB device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | + | You have chosen to repress a gene using a UVB light inducible dCas9-UVR8 & COP1-KRAB device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). |
</p> | </p> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/6/6d/6_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
- | You have chosen to repress a gene using a non inducible dCas9-G9a device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | + | You have chosen to repress a gene using a non inducible dCas9-G9a device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). |
</p> | </p> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/1/1a/7_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
- | You have chosen to repress a gene using a non inducible dCas9-KRAB device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry </a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). | + | You have chosen to repress a gene using a non inducible dCas9-KRAB device. Therefore you have to order the following plasmids from the <a id="link" href="http://parts.igem.org/Main_Page"> iGEM parts registry</a>. After receiving our plasmids, you will have to clone your target sequence into our crRNA plasmid (protocol see below). |
</p> | </p> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/3/3f/8_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> | ||
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<p> | <p> | ||
Note: <br> | Note: <br> | ||
- | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts </a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. | + | All plasmids are optimized for expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our <a id="link" href="https://2013.igem.org/Team:Freiburg/parts"> parts</a> side). This system was tested mainly in CHO-K1 and HEK-293T cells. |
</p> | </p> | ||
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</ol> | </ol> | ||
- | + | <a href="https://static.igem.org/mediawiki/2013/3/32/9_manual_pdf_freiburg.pdf"> <img src="https://static.igem.org/mediawiki/2013/9/9d/Printknopf_freiburg_13.png" width="100px" border="0" style="margin-left:350px; margin-top: 50px;" alt=""> </a> | |
</div> | </div> |
Latest revision as of 00:59, 29 October 2013
The uniCAS toolkit - Customize your experiments!
You want to have a maximum of activation or repression of your genes by a minimal effort? Then you
have to use the uniCAS toolkit provided by the iGEM team Freiburg 2013. All you have to do is:
- Click yourself through the routine below
- Order the appropriate plasmids and oligos
- Conduct a minimal of cloning
- Start your personalized experiment