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Revision as of 14:21, 9 September 2012 (view source) Davsie (Talk | contribs) ← Older edit		Revision as of 14:34, 9 September 2012 (view source) Davsie (Talk | contribs) Newer edit →
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-		+	<br><br>Because the two thymins are already in the cloning vector they are not intresting for our TAL protein:
-	<br>Because the two thymins are already in the cloning vector they are not intresting for our TAL protein:	+
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-	Two build this sequence from our toolkit we need to split it up in pairs of two:	+
		+	<br><br>To build this sequence from our toolkit we need to split it up in pairs of two:
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		+	{|align="left"
		+	|[[Image:sequence3.png|350px|no frame]]
		+	|}
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		+	<br><br>Now we need to give our pairs position numbers inside the TAL protein:
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		+	{|align="left"
		+	|[[Image:sequence4.png|500px|no frame]]
		+	|}

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Revision as of 14:34, 9 September 2012

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																					Building a Toolkit	Using the Toolkit		Experiments

Using the Toolkit

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Here we try to give you something like a short manual on how to use our toolkit to design TAL poteins.

Step 1. The Experiment

First you need to think about your experiment and what you want to do, when working with TAL proteins its pretty clear you want to target a dna sequence. To choose your sequence you need to know some of the limitations of TAL proteins to pick it the right way.

1. Every TAL protein starts and ends with a Thymin

This Thymin is already inserted in front of your TAL protein when you use our toolkit but it should also be in front of the first base of your sequence

2. Your sequence must be twelve bases long

Our Toolbox is optimized for sequences of twelve plus two (the thymin at upstream and downstrem) this lenght guarantes a high enough specifity and at the same time a librarysize that good to handle.

If you found a sequence that suits these limitations you can start building your TAL protein.

Step 1. Building a TAL

For building your TAL you start with your selected sequence, in this amnual we use a fictive sequence that you can substitute with your own.

Our sequence will be:

Because the two thymins are already in the cloning vector they are not intresting for our TAL protein:

To build this sequence from our toolkit we need to split it up in pairs of two:

Now we need to give our pairs position numbers inside the TAL protein: