Team:Heidelberg/Notebook/material
From 2008.igem.org
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+ | |||
+ | ===Kits=== | ||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=350px | Kit || width=350px | supplier | ||
+ | |-style="height:20px" | ||
+ | | || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |CompactPrep Plasmid Maxi Kit || Qiagen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |HISpeed Plasmid Maxi Kit || Qiagen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |MaxPlax™ Lambda Packaging Extracts || EPICENTRE Biotechnologies | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |QIAEX II Gel Extraction Kit || Qiagen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |QIAGEN Lambda Mini Kit || Qiagen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |QIAprep Spin Mini Kit || Qiagen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |QIAquick Gel Extraction Kit || Qiagen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |QIAquick PCR Purification Kit || Qiagen | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | ===Marker=== | ||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=350px | Marker || width=350px | supplier | ||
+ | |-style="height:20px" | ||
+ | | || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |GeneRulerTM High Range DNA Ladder || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |GeneRulerTM 1kb DNA Ladder Mix || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |GeneRulerTM 1kb Plus DNA Ladder Mix || MBI Fermentas | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | ===Enzymes=== | ||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=350px | Enzym || width=350px | supplier | ||
+ | |-style="height:20px" | ||
+ | | || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Pfu DNA polymerase || Stratagene | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Pfu turbo DNA polymerase || Stratagene | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Phusion DNA polymerase || Finnzymes | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Taq DNA polymerase || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |T4 DNA ligase || MBI Fermentas / New England Biolabs | ||
+ | |-style="height:20px" | ||
+ | | || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |AgeI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BamHI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BglI || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BseJI || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BspEI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |DpnI || Roche Diagnostics / New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |DraI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |EcoRI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |EcoRV || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |HindIII || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |KpnI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |NcoI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |NdeI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |NotI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |PstI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |SacI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |SalI || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |ScaI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |SfcI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |SmiI || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |SpeI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |SpeI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |XbaI || New England Biolabs | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |XhoI || MBI Fermentas | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |XmaI || New England Biolabs | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | ===Plasmidvectors=== | ||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=200px | Name || width=250px | application || width=250px | reference | ||
+ | |-style="height:20px" | ||
+ | | || || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_B0015 || terminator || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_F1610 || LuxI || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_I15030 || AI-1 amplifier || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_I20260 || GFP || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_J01003 || oriT || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_J16002 || cloning vector || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_J23107 || constitutive promotor || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |BBa_T9002 || AI-1 GFP receiver || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |CheY-mCherry || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |ColE1 || colicin E1 || DSMZ | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |ColE9-J || colicin E9 || C. Kleanthous, University of York | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pBad18 || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pBad33 || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pBluescript II SK (+) || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pDest15 || cloning vector || DKFZ Library | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pDK4 || visualization || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pDK48 || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pDK58 || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pDK6 || visualization || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pED374 || oriT || K. Derbyshire, Wadsworth | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pES16 || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pMMB863 || oriT || M. Bagdasarian, MSU | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pQE-30 || cloning vector || Invitrogen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pSB1A2 || cloning vector || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pSB2K3 || cloning vector || http://partsregistry.org | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pTrc99a || cloning vector || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |pUB307 || helper plasmid || E. Lanka, BfR | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |RP4 || helper plasmid || M. Bagdasarian, MSU | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | ===Synthetic oligonucleotides=== | ||
+ | |||
+ | ===Phages=== | ||
+ | |||
+ | ===Bacteria=== | ||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=200px | E.coli strain || width=250px | usage || width=250px | reference | ||
+ | |-style="height:20px" | ||
+ | | || || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |DH5a || amplification of plasmids || Invitrogen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |HCB33 || swarm assays || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |MG1655 || swarm assays || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |TOP10 || amplification of plasmids || Invitrogen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |UU1250 || chemotaxis receptor knock out || V. Sourjik, ZMBH | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | ===Bacteria Growth Media=== | ||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=200px | E.coli strain || width=250px | usage || width=250px | reference | ||
+ | |-style="height:20px" | ||
+ | | || || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |DH5a || amplification of plasmids || Invitrogen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |HCB33 || swarm assays || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |MG1655 || swarm assays || V. Sourjik, ZMBH | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |TOP10 || amplification of plasmids || Invitrogen | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |UU1250 || chemotaxis receptor knock out || V. Sourjik, ZMBH | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | |||
+ | 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: | ||
+ | |||
+ | |||
+ | {| class="wikitable" | ||
+ | |- bgcolor=grey | ||
+ | ! height=20px, width=200px | Antibiotic || width=250px | concentration | ||
+ | |-style="height:20px" | ||
+ | | || | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Ampicillin || 100 µg/ml | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Chloramphenicol || 35 µg/ml | ||
+ | |-align="center" | ||
+ | | style="font-weight:bold;" |Kanamycin || 50 µg/ml | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | The same concentrations of antibiotics were used for selection of resistant in media. |
Revision as of 14:57, 28 October 2008
Contents |
Material
Buffers & Solution
SM | 50 mM | Tris, pH 7.5 |
20 mM | MgSO4 | |
50 mM | NaCl | |
0.01 % | gelatine | |
Tethering buffer, pH 7.0 | 10 mM | potassium phosphate |
(K2HPO4 : KH2PO4 = 1 :1 ) | ||
100 µM | EDTA | |
1 µM | L-Methionine | |
10 mM | lactic acid | |
M63 salt | 3 g/l | KH2PO4 |
9 g/l | K2HPO4 | |
4 g/l | (NH4)2SO4 | |
0.5 g/l | FeSO4 | |
Amino acid mix | 5 g/l | L-threonine |
5 g/l | L-histidin | |
5 g/l | L-leucine | |
5 g/l | L-methionine | |
M9 salts | 64 g/l | Na2HPO4 x 7H2O |
15 g/l | KH2PO4 | |
2.5 g/l | NaCl | |
5 g/l | NH4Cl |
Kits
Kit | supplier |
---|---|
CompactPrep Plasmid Maxi Kit | Qiagen |
HISpeed Plasmid Maxi Kit | Qiagen |
MaxPlax™ Lambda Packaging Extracts | EPICENTRE Biotechnologies |
QIAEX II Gel Extraction Kit | Qiagen |
QIAGEN Lambda Mini Kit | Qiagen |
QIAprep Spin Mini Kit | Qiagen |
QIAquick Gel Extraction Kit | Qiagen |
QIAquick PCR Purification Kit | Qiagen |
Marker
Marker | supplier |
---|---|
GeneRulerTM High Range DNA Ladder | MBI Fermentas |
GeneRulerTM 1kb DNA Ladder Mix | MBI Fermentas |
GeneRulerTM 1kb Plus DNA Ladder Mix | MBI Fermentas |
Enzymes
Enzym | supplier |
---|---|
Pfu DNA polymerase | Stratagene |
Pfu turbo DNA polymerase | Stratagene |
Phusion DNA polymerase | Finnzymes |
Taq DNA polymerase | MBI Fermentas |
T4 DNA ligase | MBI Fermentas / New England Biolabs |
AgeI | New England Biolabs |
BamHI | New England Biolabs |
BglI | MBI Fermentas |
BseJI | MBI Fermentas |
BspEI | New England Biolabs |
DpnI | Roche Diagnostics / New England Biolabs |
DraI | New England Biolabs |
EcoRI | New England Biolabs |
EcoRV | New England Biolabs |
HindIII | New England Biolabs |
KpnI | New England Biolabs |
NcoI | New England Biolabs |
NdeI | New England Biolabs |
NotI | New England Biolabs |
PstI | New England Biolabs |
SacI | New England Biolabs |
SalI | MBI Fermentas |
ScaI | New England Biolabs |
SfcI | New England Biolabs |
SmiI | MBI Fermentas |
SpeI | New England Biolabs |
SpeI | New England Biolabs |
XbaI | New England Biolabs |
XhoI | MBI Fermentas |
XmaI | New England Biolabs |
Plasmidvectors
Name | application | reference |
---|---|---|
BBa_B0015 | terminator | http://partsregistry.org |
BBa_F1610 | LuxI | http://partsregistry.org |
BBa_I15030 | AI-1 amplifier | http://partsregistry.org |
BBa_I20260 | GFP | http://partsregistry.org |
BBa_J01003 | oriT | http://partsregistry.org |
BBa_J16002 | cloning vector | http://partsregistry.org |
BBa_J23107 | constitutive promotor | http://partsregistry.org |
BBa_T9002 | AI-1 GFP receiver | http://partsregistry.org |
CheY-mCherry | cloning vector | V. Sourjik, ZMBH |
ColE1 | colicin E1 | DSMZ |
ColE9-J | colicin E9 | C. Kleanthous, University of York |
pBad18 | cloning vector | V. Sourjik, ZMBH |
pBad33 | cloning vector | V. Sourjik, ZMBH |
pBluescript II SK (+) | cloning vector | V. Sourjik, ZMBH |
pDest15 | cloning vector | DKFZ Library |
pDK4 | visualization | V. Sourjik, ZMBH |
pDK48 | cloning vector | V. Sourjik, ZMBH |
pDK58 | cloning vector | V. Sourjik, ZMBH |
pDK6 | visualization | V. Sourjik, ZMBH |
pED374 | oriT | K. Derbyshire, Wadsworth |
pES16 | cloning vector | V. Sourjik, ZMBH |
pMMB863 | oriT | M. Bagdasarian, MSU |
pQE-30 | cloning vector | Invitrogen |
pSB1A2 | cloning vector | http://partsregistry.org |
pSB2K3 | cloning vector | http://partsregistry.org |
pTrc99a | cloning vector | V. Sourjik, ZMBH |
pUB307 | helper plasmid | E. Lanka, BfR |
RP4 | helper plasmid | M. Bagdasarian, MSU |
Synthetic oligonucleotides
Phages
Bacteria
E.coli strain | usage | reference |
---|---|---|
DH5a | amplification of plasmids | Invitrogen |
HCB33 | swarm assays | V. Sourjik, ZMBH |
MG1655 | swarm assays | V. Sourjik, ZMBH |
TOP10 | amplification of plasmids | Invitrogen |
UU1250 | chemotaxis receptor knock out | V. Sourjik, ZMBH |
Bacteria Growth Media
E.coli strain | usage | reference |
---|---|---|
DH5a | amplification of plasmids | Invitrogen |
HCB33 | swarm assays | V. Sourjik, ZMBH |
MG1655 | swarm assays | V. Sourjik, ZMBH |
TOP10 | amplification of plasmids | Invitrogen |
UU1250 | chemotaxis receptor knock out | V. Sourjik, ZMBH |
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 |
---|---|
Ampicillin | 100 µg/ml |
Chloramphenicol | 35 µg/ml |
Kanamycin | 50 µg/ml |
The same concentrations of antibiotics were used for selection of resistant in media.