Difference between revisions of "Team:ETH Zurich/Materials"
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<li>Novaspec Plus™ Visible Spectrophotometer</li> | <li>Novaspec Plus™ Visible Spectrophotometer</li> | ||
<a href="http://www.gelifesciences.com/webapp/wcs/stores/servlet/ProductDisplay?categoryId=11012&catalogId=10101&productId=20430&storeId=11782&langId=-1"><p>for further information click here</p></a> | <a href="http://www.gelifesciences.com/webapp/wcs/stores/servlet/ProductDisplay?categoryId=11012&catalogId=10101&productId=20430&storeId=11782&langId=-1"><p>for further information click here</p></a> | ||
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<li>MettlerToledo XS4002S DeltaRange® Balance</li> | <li>MettlerToledo XS4002S DeltaRange® Balance</li> | ||
<a href="http://us.mt.com/us/en/home/products/Laboratory_Weighing_Solutions/Precision_Balances/XS_Precision_Balances/XS_4002_S_Dual_Range.html"><p>for further information click here</p></a> | <a href="http://us.mt.com/us/en/home/products/Laboratory_Weighing_Solutions/Precision_Balances/XS_Precision_Balances/XS_4002_S_Dual_Range.html"><p>for further information click here</p></a> | ||
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<li>Helixx Technologies MyRun®</li> | <li>Helixx Technologies MyRun®</li> | ||
<a href="http://www.helixxtec.com/helixx/c/iMyRun/iMyRun.htm"><p>for further information click here</p></a> | <a href="http://www.helixxtec.com/helixx/c/iMyRun/iMyRun.htm"><p>for further information click here</p></a> | ||
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<li>Eppendorf Thermomixer compact</li> | <li>Eppendorf Thermomixer compact</li> | ||
<a href="http://www.eppendorf.com/int/index.php?pb=93a83e2a7b7b1b05&action=products&contentid=1&catalognode=9840"><p>for further information click here</p></a> | <a href="http://www.eppendorf.com/int/index.php?pb=93a83e2a7b7b1b05&action=products&contentid=1&catalognode=9840"><p>for further information click here</p></a> | ||
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<li>BD LSRFortessa™ cell analyzer</li> | <li>BD LSRFortessa™ cell analyzer</li> | ||
<a href="http://www.bdbiosciences.com/us/instruments/research/cell-analyzers/bd-lsrfortessa/m/751752?cc=US"><p>for further information click here</p></a> | <a href="http://www.bdbiosciences.com/us/instruments/research/cell-analyzers/bd-lsrfortessa/m/751752?cc=US"><p>for further information click here</p></a> | ||
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<li>ODYSSEY® CLx Infrared Imaging System</li> | <li>ODYSSEY® CLx Infrared Imaging System</li> | ||
<a href="http://www.licor.com/bio/products/imaging_systems/odyssey/"><p>for further information click here</p></a> | <a href="http://www.licor.com/bio/products/imaging_systems/odyssey/"><p>for further information click here</p></a> | ||
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<li>Thermo Scientific Heraeus Incubator</li> | <li>Thermo Scientific Heraeus Incubator</li> | ||
<a href="https://webshop.fishersci.com/insight2_uk/getProduct.do?productCode=11626250&resultSetPosition=0"><p>for further information click here</p></a> | <a href="https://webshop.fishersci.com/insight2_uk/getProduct.do?productCode=11626250&resultSetPosition=0"><p>for further information click here</p></a> | ||
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<li>Eppendorf Centrifuge 5810 R</li> | <li>Eppendorf Centrifuge 5810 R</li> | ||
<a href="http://www.eppendorf.com/int/index.php?pb=01edea0fe7a0e358&action=products&contentid=1&catalognode=10019"><p>for further information click here</p></a> | <a href="http://www.eppendorf.com/int/index.php?pb=01edea0fe7a0e358&action=products&contentid=1&catalognode=10019"><p>for further information click here</p></a> | ||
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<li>Thermo Scientific NanoDrop® 2000 UV-Vis spectrophotometer combined with NanoDrop 2000 / 2000c Software</li> | <li>Thermo Scientific NanoDrop® 2000 UV-Vis spectrophotometer combined with NanoDrop 2000 / 2000c Software</li> | ||
<a href="http://www.nanodrop.com/Productnd2000overview.aspx"><p>for further information click here</p></a> | <a href="http://www.nanodrop.com/Productnd2000overview.aspx"><p>for further information click here</p></a> | ||
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<li>Applied Biosystems Veriti 96Well Thermal Cycler</li> | <li>Applied Biosystems Veriti 96Well Thermal Cycler</li> | ||
<a href="https://www.thermofisher.com/ch/en/home/life-science/pcr/thermal-cyclers-realtime-instruments/thermal-cyclers/veriti-thermal-cycler.html"><p>for further information click here</p></a> | <a href="https://www.thermofisher.com/ch/en/home/life-science/pcr/thermal-cyclers-realtime-instruments/thermal-cyclers/veriti-thermal-cycler.html"><p>for further information click here</p></a> | ||
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<a href="http://www.tecan.com/platform/apps/product/index.asp?MenuID=1812&ID=1916&Menu=1&Item=21.2.10.1"><p>for further information (platereader) click here</p></a> | <a href="http://www.tecan.com/platform/apps/product/index.asp?MenuID=1812&ID=1916&Menu=1&Item=21.2.10.1"><p>for further information (platereader) click here</p></a> | ||
<a href="http://www.tecan.ch/platform/apps/product/index.asp?MenuID=1817&ID=1924&Menu=1&Item=21.7.8"><p>for further information (software) click here</p></a> | <a href="http://www.tecan.ch/platform/apps/product/index.asp?MenuID=1817&ID=1924&Menu=1&Item=21.7.8"><p>for further information (software) click here</p></a> | ||
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<li>Kuhner Climo-Shaker ISF1-X</li> | <li>Kuhner Climo-Shaker ISF1-X</li> | ||
<a href="http://www.kuhner.com/de/produkte/schuettelmaschinen/inkubatoren/isf1-x.html"><p>for further information click here</p></a> | <a href="http://www.kuhner.com/de/produkte/schuettelmaschinen/inkubatoren/isf1-x.html"><p>for further information click here</p></a> | ||
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<li>Eppendorf Centrifuge 5424 R</li> | <li>Eppendorf Centrifuge 5424 R</li> | ||
<a href="http://www.labdepotinc.com/Product_Details~id~450~pid~59833.aspx"><p>for further information click here</p></a> | <a href="http://www.labdepotinc.com/Product_Details~id~450~pid~59833.aspx"><p>for further information click here</p></a> | ||
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<a href="http://www.biostep.de/i18n_en_DE/products/Bio_Imaging_components_1776/Tripod__dark_hoods_and_accessories_1863/Dark_hoods_1865/Dark_Hood_DH_40_with_control_panel_and_display_6/index.html"><p>for further information (hood) click here</p></a> | <a href="http://www.biostep.de/i18n_en_DE/products/Bio_Imaging_components_1776/Tripod__dark_hoods_and_accessories_1863/Dark_hoods_1865/Dark_Hood_DH_40_with_control_panel_and_display_6/index.html"><p>for further information (hood) click here</p></a> | ||
<a href="https://www.biostep.de/products/Bio_Imaging_software_1843/Acquisition_and_control_software_1844/argus_X1___for_cameras_and_scanners_1845/biostep_argusX1__basic_licence_1/index.html"><p>for further information (software) click here</p></a> | <a href="https://www.biostep.de/products/Bio_Imaging_software_1843/Acquisition_and_control_software_1844/argus_X1___for_cameras_and_scanners_1845/biostep_argusX1__basic_licence_1/index.html"><p>for further information (software) click here</p></a> | ||
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<li>Scientific Industries VortexGenie®2</li> | <li>Scientific Industries VortexGenie®2</li> | ||
<a href="http://www.scientificindustries.com/genie2.html"><p>for further information click here</p></a> | <a href="http://www.scientificindustries.com/genie2.html"><p>for further information click here</p></a> | ||
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Revision as of 09:31, 18 September 2015
- Project
- Modeling
- Lab
- Human
Practices - Parts
- About Us
Materials
Bacterial strains and Mammalian Cell Lines
TOP10
The bacterial strain we used as a base for all our transformations was TOP10. These cells were provided by the Panke Group at ETH Zurich.
A list of all strains we generated in the course of our project can be found here (link).
Jurkat Cells
Immortalized human T lymphocyte cell line. We used this cell line as a model line for cancer cells with elevated lactate production and TRAIL susceptibility (papers). A big thank you to the Tay Group at ETH Zurich who kindly offered us an aliquot of their cells.
HL60
Cell line derived from acute myeloid leukemia. We used this cell line as a model line for cancer cells with elevated lactate production and TRAIL susceptibility (papers). Many thanks to the Neri Group at ETH Zurich who kindly offered us an aliquot of their cells.
SK-BR-3
HER2-positive breast cancer cell line. We used this cell line to validate the chimeric receptors of the Stockholm iGEM team. We are very grateful to Michal Stanczak from the University Hospital of Basel for offering us a sample of this cell line.
3T3
Murine fibroblast cell line. We used this cell line as a model line for healthy cells which are restistant to TRAIL and produce normal levels of lactate. Many thanks to the Tay Group at ETH Zurich who kindly offered us an aliquot of their cells.
3T3 P65 -/-
Murine fluorescent fibroblast cell line.
- H2B-GFP: nuclear staining
- P65- dsRed: cytoskeleton
Buffers and media
Annexin V binding buffer
- 10 mM HEPES
- 140 mM NaCl
- 2.5 mM
- 250 mM CaCl2
- pH 7.4
- Filter the solution through a 0.2 µm filter.
Jurkat medium
- 2 mL MEM 1x
- 20 mL FBS 100%
- 2 mL GlutaMax 100x
- 0.4 mL Penicillin (10k units/mL) /Streptomyocin (10k µg/mL )
- 200 mL RPMI
Terrific Broth
- 2.32 g/L KH2PO4
- 12.54 g/L K2HPO4
- 1.2% Tryptone
- 2.4% Yeast Extract
- 0.4% Gylcerol
- Filter the solution through a 0.2 µm filter.
SOC medium
- 20 g/L Tryptophane
- 5 g/L Yeast extract
- 0.5 g/L NaCl
- 250 mM KCl
- 1M MgCl2
- 50% (w/v) sterile glucose
- Filter the solution through a 0.2 µm filter.
TFB1
- 100 mM RbCl
- 50 mM MnCl2
- 30 mM potassium acetate
- 10 mM CaCl2
- 15%glycerol
TFB2
- 100 mM MOPS
- 50 mM RbCl
- 75 mM CaCl2
- 15%glycerol
3T3 medium
- 500 mL D-MEM 1x
- 50 mL FBS 100%
- 5 mL GlutaMax 100x
- 5 mL Penicillin (10k units/mL) /Streptomyocin (10k µg/mL )
Chemicals
LB-Broth
- Product name: BD Difco™ LB Broth, Miller (Luria-Bertani)
- Company: BD
- REF: 244620
for further information click here
Agarose
- Product name: Agarose
- Company: Sigma-Aldrich
- REF: A9539-500G
for further information click here
Agar
- Product name: Agar für die Bakteriologie BC
- Company: Sigma-Aldrich
- REF: A0949
for further information click here
Glycerol
- Product: Glycerol anhydrous BioChemica BC
- Company: AppliChem
- REF: A1123
for further information click here
2-log DNA ladder
- Product: 2-Log DNA Ladder (0.1-10.0 kb)
- Company: New England BioLabs
- REF: N3200S
for further information click here
Taq DNA Ligase
- Product: Taq DNA Ligase
- Company: New England BioLabs
- REF: M0208S
for further information click here
Cutsmart Buffer
- Product: CutSmart® Buffer
- Company: New England BioLabs
- REF: B7204S
for further information click here
EcoRI
- Product: EcoRI-HF®
- Company: New England BioLabs
- REF: R3101S
for further information click here
BsaI-HF
- Product: BsaI-HF®
- Company: New England BioLabs
- REF: R3535S
for further information click here
Q5 Master Mix
- Product: Q5® High-Fidelity 2X Master Mix
- Company: New England BioLabs
- REF: M0492S
for further information click here
dNTP Mix
- Product: Deoxynucleotide (dNTP) Solution Mix
- Company: New England BioLabs
- REF: N1201A
for further information click here
Taq DNA Polymerase
- Product: Taq DNA Polymerase with ThermoPol® Buffer
- Company: New England BioLabs
- REF: M0276L
for further information click here
AnnexinV-Alexa
- Product: Annexin V, Alexa Fluor® 488 conjugate
- Company: ThermoFischer Scientific
- REF: A13201
for further information click here
Protein G coated Beads
- Product: Pierce™ Protein G Magnetic Beads
- Company: Thermo Fischer Scientific
- REF: 88847
for further information click here
Streptavidin Beads
- Product: Roti®-MagBeads Streptavidin
- Company: Carl Roth
- REF: HP57.2
for further information click here
Red Gel Stain
- Product: GelRed Nucleic Acid Gel Stain, 10,000X in water
- Company: Biotium
- REF: 41003
for further information click here
Anti-Human-AnnexinV
- Product: anti-human Annexin V/Biotin
- Company: Affymetrix eBioscience
- REF: BMS147
for further information click here
Gel Loading Dye
- Product: Gel Loading Dye, Purple (6X), no SDS
- Company: New England BioLabs
- REF: B7025S
for further information click here
T4 DNA Ligase
- Product: T4 DNA Ligase
- Company: New England BioLabs
- REF: M0202S
for further information click here
DNA Ligase Buffer
- Product: Taq DNA Ligase Reaction Buffer
- Company: New England BioLabs
- REF: B0208S
for further information click here
DMEM
- Product: DMEM, high glucose, no glutamine, no phenol red
- Company: Gibco ThermoFisher Scientific
- REF: 31053-028
for further information click here
DMEM with Glutamine
- Product: DMEM cell culture media
- Company: Gibco ThermoFisher Scientific
- REF: 41965-039
for further information click here
RPMI
- Product: RPMI 1640 Medium, no phenol red
- Company: Gibco ThermoFisher Scientific
- REF: 11835-063
for further information click here
Accutase
- Product: StemPro® Accutase® Cell Dissociation Reagent
- Company: Gibco ThermoFisher Scientific
- REF: A11105-01
for further information click here
SuperKillerTRAIL™
- Product: SuperKillerTRAIL™
- Company: Enzo
- REF: N3200SALX-201-115-C010
for further information click here
TRAIL
- Product: TRAIL / APO2 Ligand Human Recombinant
- Company: ProspecBio
- REF: CYT-443
for further information click here
Kits
Plasmid Purification
- Product: ZR Plasmid Miniprep ™-Classic
- Company: Zymo Research
- Lot: ZRC182892
- Catalog No: D4016
Changes to the protocol included an extra step to dry the column before elution and the use of water for elution instead of elution buffer. For elution of large plasmids we used prewarmed water.
Gel Extraction
- Product: ZymocleanTM Gel DNA Recovery Kit
- Company: Zymo Research
- Lot: ZRC183055
- Catalog No: D4002
Changes to the protocol included an extra step to dry the column before elution and the use of water for elution instead of elution buffer. For elution of large plasmids we used prewarmed water.
Lactate Detection Kit
- Product: L-Lactic Acid
- Company: Megazyme
- Lot:
- Catalog No:
Changes to the protocol included an extra step to dry the column before elution and the use of water for elution instead of elution buffer. For elution of large plasmids we used prewarmed water.
The detection of lactate using this kit relies on two enzymatic reactions. In the first, L-lactate and NAD&sup+; were converted into pyruvate and NADH+H&sup+; by L-lactate dehydrogenase. In the coupled reaction, pyruvate is used with D-glutamate by the D-glutamate-pyruvate transaminase to have the equilibrium of the first reaction in favour of the pyruvate. The concentration of NADH is finally measured as the optical density at 340 nm (OD340).
Machines
Emulsifier
- EmulsiFlex-C3 from AVESTIN, Inc. and AVESTIN Europe GmbH/li>
for further information click here
Fluorescence Scanner for Western Blots
- ODYSSEY® CLx Infrared Imaging System
for further information click here
Nano Drop
- Thermo Scientific NanoDrop® 2000 UV-Vis spectrophotometer combined with NanoDrop 2000 / 2000c Software
for further information click here
Plate Reader
- Tecan Infinite M200 Pro™ combined with Tecan iControl TM software
for further information (platereader) click here
for further information (software) click here
UV- and Dark Hood
- Biostep Dark-Hood DH-50™ combined with Argus-X1™ software and Biostep argusX1™ basic licence
for further information (hood) click here
for further information (software) click here
We would like to thank our sponsors
