Team:Sharon MA Aquila/Project

From 2012hs.igem.org

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**Electroporation Buffer:(0.4 M sucrose, 1 mM MgCl2, 5 mM Kh2PO4; PH 6)
**Electroporation Buffer:(0.4 M sucrose, 1 mM MgCl2, 5 mM Kh2PO4; PH 6)
**Milk Medium: (0.2 M sucrose, 5% skim milk, 0.1% yeast extract, 1% Casamino Acids, 25mM MgCl2)
**Milk Medium: (0.2 M sucrose, 5% skim milk, 0.1% yeast extract, 1% Casamino Acids, 25mM MgCl2)
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**Erythromycin, Chloramphenicol, or other antibiotics (for selection)
+
**Erythromycin (for selection)
**Gene Pulser and Pulse Controller Electrophoration apparatus
**Gene Pulser and Pulse Controller Electrophoration apparatus
**Gaspak or method to generate anaerobic conditions
**Gaspak or method to generate anaerobic conditions

Revision as of 19:32, 3 April 2012


Parts

  • [http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=11842&Template=bacteria Lactobacillus delbrueckii subsp. bulgaricus] (from ATCC)
  • [http://partsregistry.org/Part:BBa_K652001 RiAFP Part] (from the Parts Registry BBa_K652001)
  • [http://partsregistry.org/Part:BBa_K128006 LacS Promoter] (from the Parts Registry BBa_K128006)
  • [http://partsregistry.org/Part:BBa_K128007 Plasmid] to confer Erythromycin resistance to bacteria

Materials

  • Culturing:
    • [http://www.bd.com/ds/productCenter/288110.asp MRS broth]
    • [http://www.bd.com/ds/productCenter/288210.asp MRS agar]
  • Bacterial Transformation:
    • Electroporation Buffer:(0.4 M sucrose, 1 mM MgCl2, 5 mM Kh2PO4; PH 6)
    • Milk Medium: (0.2 M sucrose, 5% skim milk, 0.1% yeast extract, 1% Casamino Acids, 25mM MgCl2)
    • Erythromycin (for selection)
    • Gene Pulser and Pulse Controller Electrophoration apparatus
    • Gaspak or method to generate anaerobic conditions
  • Miniprep
    • TES(50mM Tris[pH 7]
    • 20mM EDTA, 0.2 mM sucrose)
    • 10mg/ml lysozyme
    • 70U/ml mutanolysin


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