Melbourne/Lab BL Notebook
From 2007.igem.org
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==Aug 2007: Primer design for PCR stitching== | ==Aug 2007: Primer design for PCR stitching== | ||
In order to stitch Nt-SopII to ComP at the sites detailed [[Melbourne/Blue_Photosensor_Background#Chosen_Fusion_Sites|here]], [[Melbourne/Lab BL Notebook/PrimersI|these primers]] were designed. We aimed to have at least 18bp on each half of the stitch, while adhering to the standard primer design guidelines. | In order to stitch Nt-SopII to ComP at the sites detailed [[Melbourne/Blue_Photosensor_Background#Chosen_Fusion_Sites|here]], [[Melbourne/Lab BL Notebook/PrimersI|these primers]] were designed. We aimed to have at least 18bp on each half of the stitch, while adhering to the standard primer design guidelines. | ||
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Revision as of 12:13, 10 October 2007
<Return to Lab notebook> <team home page>
Aug 2007: Order ComP and ComA from GENEART
We were counting on [http://partsregistry.org/Part:BBa_J51000 ComP] and [http://partsregistry.org/Part:BBa_J51001 ComA] being available in the 2007 iGEM distribution. As this was not the case, and we had DNA synthesis bases to burn, we ended up ordering them from GENEART. The original sequences, as described in the above links, were optimized for expression in e.coli and all cis-acting sites and assembly restriction sites were removed. We also included biobrick prefix/suffix and vf2 and vr primer binding sites. The two sequences, now labeled ComA' and ComP' took around 3 weeks after confirmation of order to be dispatched. The sequences have been uploaded below:
ComA' sequence (As ordered sequence)
ComP' sequence (As ordered sequence)
Aug 2007: Primer design for PCR stitching
In order to stitch Nt-SopII to ComP at the sites detailed here, these primers were designed. We aimed to have at least 18bp on each half of the stitch, while adhering to the standard primer design guidelines.
