Team:LMU-Munich/Lab Notebook/Protocols

From 2012.igem.org

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==Protocols==
 
On this page, we offer our unique protocols to the public.
On this page, we offer our unique protocols to the public.
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PCR protocol for long flanking homology PCR. You might need those PCR products for interrupting genes with resistance cassettes.
PCR protocol for long flanking homology PCR. You might need those PCR products for interrupting genes with resistance cassettes.
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[https://static.igem.org/mediawiki/2012/6/67/LMU-Munich_2012_Removal_of_germination_genes.pdf Example: Using LFH PCR to replace germination genes with resistance cassettes]
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An explanation of how we used this protocol for how we knocked out our germination genes using LFH.
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9. Mikroskopie der Sporen
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9. [https://static.igem.org/mediawiki/2012/e/e9/LMU-Munich_2012_Protocol_for_enhancement_of_mature_spore_numbers.pdf Enhancement of mature spore numbers]
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Protocol to enrich the spore numbers and purification from vegetative cells.
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11. Plate reader assay (can also be used with E. coli cells)
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11. [https://static.igem.org/mediawiki/2012/e/e6/LMU-Munich_2012_Protocol_Plate_Reader.pdf Luminescence measurement]
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This protocol explains how cultures were treated in advance of a plate reader assay.
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E. coli antibiotics concentrations
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[http://partsregistry.org/Help:Protocols/Competent_Cells Competent Cells]
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''E.coli'' competent cells and transformation. We used the [http://ecoliwiki.net/colipedia/index.php/XL-1_Blue XL1 blue strain].
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E.coli trafo
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[https://static.igem.org/mediawiki/2012/1/1f/LMU-Munich_2012_Alkaline_Lysis_Plasmid_Preparation.pdf Alkaline Lysis Plasmid Preparation]
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Alkaline Lysis Plasmid Preparation for ''E. coli''
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E.coli dreck-prep
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Ligation, Verdau
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[http://openwetware.org/wiki/Silver:_Restriction_Digest Restriction digest]
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[http://openwetware.org/wiki/Silver:_Ligation Ligation]
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Standard restriction digest and ligation. We often used longer incubation times (up to over night) and did not dephosphorylate the backbones, if they had incompatible sticky ends.
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Kompetente E.colis
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lacZ assay in E.coli
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[https://static.igem.org/mediawiki/2012/b/bb/LMU-Munich_2012_%C3%9F-Galactosidase_Assay_E._coli.pdf β-Galactosidase Assay in ''E. coli'']
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Quantitative ''lacZ''-assay in ''E.coli''.
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(Proteinase K)
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(Western Blot)
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Latest revision as of 02:00, 27 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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