From 2008.igem.org
Material
Buffers & Solution
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| SM | 50 mM | Tris, pH 7.5
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| | 20 mM | MgSO4
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| | 50 mM | NaCl
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| | 0.01 % | gelatine
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| | |
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| Tethering buffer, pH 7.0 | 10 mM | potassium phosphate
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| | | (K2HPO4 : KH2PO4 = 1 :1 )
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| | 100 µM | EDTA
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| | 1 µM | L-Methionine
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| | 10 mM | lactic acid
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| M63 salt | 3 g/l | KH2PO4
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| | 9 g/l | K2HPO4
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| | 4 g/l | (NH4)2SO4
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| | 0.5 g/l | FeSO4
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| Amino acid mix | 5 g/l | L-threonine
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| | 5 g/l | L-histidin
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| | 5 g/l | L-leucine
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| | 5 g/l | L-methionine
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| | |
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| M9 salts | 64 g/l | Na2HPO4 x 7H2O
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| | 15 g/l | KH2PO4
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| | 2.5 g/l | NaCl
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| | 5 g/l | NH4Cl
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Kits
| Kit | supplier
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| CompactPrep Plasmid Maxi Kit | Qiagen
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| HISpeed Plasmid Maxi Kit | Qiagen
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| MaxPlax™ Lambda Packaging Extracts | EPICENTRE Biotechnologies
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| QIAEX II Gel Extraction Kit | Qiagen
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| QIAGEN Lambda Mini Kit | Qiagen
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| QIAprep Spin Mini Kit | Qiagen
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| QIAquick Gel Extraction Kit | Qiagen
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| QIAquick PCR Purification Kit | Qiagen
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Marker
| Marker | supplier
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| GeneRulerTM High Range DNA Ladder | MBI Fermentas
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| GeneRulerTM 1kb DNA Ladder Mix | MBI Fermentas
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| GeneRulerTM 1kb Plus DNA Ladder Mix | MBI Fermentas
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Enzymes
| Enzym | supplier
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| Pfu DNA polymerase | Stratagene
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| Pfu turbo DNA polymerase | Stratagene
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| Phusion DNA polymerase | Finnzymes
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| Taq DNA polymerase | MBI Fermentas
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| T4 DNA ligase | MBI Fermentas / New England Biolabs
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| AgeI | New England Biolabs
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| BamHI | New England Biolabs
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| BglI | MBI Fermentas
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| BseJI | MBI Fermentas
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| BspEI | New England Biolabs
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| DpnI | Roche Diagnostics / New England Biolabs
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| DraI | New England Biolabs
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| EcoRI | New England Biolabs
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| EcoRV | New England Biolabs
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| HindIII | New England Biolabs
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| KpnI | New England Biolabs
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| NcoI | New England Biolabs
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| NdeI | New England Biolabs
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| NotI | New England Biolabs
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| PstI | New England Biolabs
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| SacI | New England Biolabs
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| SalI | MBI Fermentas
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| ScaI | New England Biolabs
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| SfcI | New England Biolabs
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| SmiI | MBI Fermentas
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| SpeI | New England Biolabs
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| SpeI | New England Biolabs
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| XbaI | New England Biolabs
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| XhoI | MBI Fermentas
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| XmaI | New England Biolabs
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Plasmidvectors
| Name | application | reference
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| BBa_B0015 | terminator | http://partsregistry.org
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| BBa_F1610 | LuxI | http://partsregistry.org
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| BBa_I15030 | AI-1 amplifier | http://partsregistry.org
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| BBa_I20260 | GFP | http://partsregistry.org
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| BBa_J01003 | oriT | http://partsregistry.org
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| BBa_J16002 | cloning vector | http://partsregistry.org
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| BBa_J23107 | constitutive promotor | http://partsregistry.org
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| BBa_T9002 | AI-1 GFP receiver | http://partsregistry.org
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| CheY-mCherry | cloning vector | V. Sourjik, ZMBH
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| ColE1 | colicin E1 | DSMZ
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| ColE9-J | colicin E9 | C. Kleanthous, University of York
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| pBad18 | cloning vector | V. Sourjik, ZMBH
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| pBad33 | cloning vector | V. Sourjik, ZMBH
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| pBluescript II SK (+) | cloning vector | V. Sourjik, ZMBH
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| pDest15 | cloning vector | DKFZ Library
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| pDK4 | visualization | V. Sourjik, ZMBH
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| pDK48 | cloning vector | V. Sourjik, ZMBH
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| pDK58 | cloning vector | V. Sourjik, ZMBH
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| pDK6 | visualization | V. Sourjik, ZMBH
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| pED374 | oriT | K. Derbyshire, Wadsworth
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| pES16 | cloning vector | V. Sourjik, ZMBH
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| pMMB863 | oriT | M. Bagdasarian, MSU
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| pQE-30 | cloning vector | Invitrogen
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| pSB1A2 | cloning vector | http://partsregistry.org
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| pSB2K3 | cloning vector | http://partsregistry.org
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| pTrc99a | cloning vector | V. Sourjik, ZMBH
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| pUB307 | helper plasmid | E. Lanka, BfR
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| RP4 | helper plasmid | M. Bagdasarian, MSU
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Synthetic oligonucleotides
Phages
Bacteria
| E.coli strain | usage | reference
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| | |
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| DH5a | amplification of plasmids | Invitrogen
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| HCB33 | swarm assays | V. Sourjik, ZMBH
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| MG1655 | swarm assays | V. Sourjik, ZMBH
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| TOP10 | amplification of plasmids | Invitrogen
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| UU1250 | chemotaxis receptor knock out | V. Sourjik, ZMBH
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Bacteria Growth Media
| E.coli strain | usage | reference
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| DH5a | amplification of plasmids | Invitrogen
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| HCB33 | swarm assays | V. Sourjik, ZMBH
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| MG1655 | swarm assays | V. Sourjik, ZMBH
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| TOP10 | amplification of plasmids | Invitrogen
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| UU1250 | chemotaxis receptor knock out | V. Sourjik, ZMBH
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For cultivation of phages the media were supplied with 0.2 % maltose and 10 mM MgSO4. For preparation of agar plates 15 g/l agar, for preparation of top agar 7 g/l agar was added prior the autoclaving. For selection of resistant bacteria following antibiotics were added during cooling of at agar at about 50 °C:
| Antibiotic | concentration
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| Ampicillin | 100 µg/ml
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| Chloramphenicol | 35 µg/ml
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| Kanamycin | 50 µg/ml
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The same concentrations of antibiotics were used for selection of resistant in media.
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