Difference between revisions of "Team:ETH Zurich/Materials"
| Line 107: | Line 107: | ||
<li>Company: BD</li> | <li>Company: BD</li> | ||
<li>REF: 244620</li> | <li>REF: 244620</li> | ||
| − | + | <a href="http://catalog.bd.com/nexus-ecat/getProductDetail?productId=244620&parentCategory=1211&parentCategoryName=Microbiology%20Testing&categoryId=1269&categoryName=Dehydrated%20Culture%20Media&searchUrl="><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 121: | Line 121: | ||
<li>Company: Sigma-Aldrich</li> | <li>Company: Sigma-Aldrich</li> | ||
<li>REF: A9539-500G</li> | <li>REF: A9539-500G</li> | ||
| − | + | <a href="http://www.sigmaaldrich.com/catalog/product/sigma/a9539?lang=de®ion=CH"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 135: | Line 135: | ||
<li>Company: Sigma-Aldrich</li> | <li>Company: Sigma-Aldrich</li> | ||
<li>REF: A0949</li> | <li>REF: A0949</li> | ||
| − | + | <a href="https://www.applichem.com/shop/produktdetail/as/agar-fuer-die-bakteriologie/"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 149: | Line 149: | ||
<li>Company: AppliChem</li> | <li>Company: AppliChem</li> | ||
<li>REF: A1123</li> | <li>REF: A1123</li> | ||
| − | + | <a href="https://www.applichem.com/en/shop/product-detail/as/glycerin-wasserfrei-ibiochemicai/"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 164: | Line 164: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: N3200S</li> | <li>REF: N3200S</li> | ||
| − | + | <a href="https://www.neb.com/products/n3200-2-log-dna-ladder-01-100-kb"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 179: | Line 179: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: M0208S</li> | <li>REF: M0208S</li> | ||
| − | + | <a href="https://www.neb.com/products/m0208-taq-dna-ligase"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 193: | Line 193: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: B7204S</li> | <li>REF: B7204S</li> | ||
| − | + | <a href="https://www.neb.com/products/b7204-cutsmart-buffer"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
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<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: R3101S</li> | <li>REF: R3101S</li> | ||
| − | + | <a href="https://www.neb.com/products/r3101-ecori-hf"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 222: | Line 222: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: R0176S</li> | <li>REF: R0176S</li> | ||
| − | + | <a href="https://www.neb.com/products/r0176-dpni"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 236: | Line 236: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: M0492S</li> | <li>REF: M0492S</li> | ||
| − | + | <a href="https://www.neb.com/products/m0492-q5-high-fidelity-2x-master-mix"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 250: | Line 250: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: N1201A</li> | <li>REF: N1201A</li> | ||
| − | + | <a href="https://www.neb.com/~/media/Catalog/All-Products/690D917CFEBC46C689678BC243668395/MSDS/sdsN1201-cae.pdf"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 264: | Line 264: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: M0276L</li> | <li>REF: M0276L</li> | ||
| − | + | <a href="https://www.neb.com/products/m0267-taq-dna-polymerase-with-thermopol-buffer"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 278: | Line 278: | ||
<li>Company: ThermoFischer Scientific</li> | <li>Company: ThermoFischer Scientific</li> | ||
<li>REF: A13201</li> | <li>REF: A13201</li> | ||
| − | + | <a href="https://www.thermofisher.com/order/catalog/product/A13201"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
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<li>Company: peqlab</li> | <li>Company: peqlab</li> | ||
<li>REF: 37-5010</li> | <li>REF: 37-5010</li> | ||
| − | + | <a href="http://shop.axonlab.ch/D/include/ShowPDFDocument.asp?SessionId=700C97586C134BCEA1C5C30FBD553A03&adilng=D&ArtNr=37-5010&Nummer=3"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 306: | Line 306: | ||
<li>Company: Carl Roth</li> | <li>Company: Carl Roth</li> | ||
<li>REF: </li> | <li>REF: </li> | ||
| − | + | <a href="https://www.carlroth.com/de/de/Chemikalien/A-Z-Chemikalien/R/Roti%3Csup%3E%C2%AE%3C-sup%3E-MagBeads/Roti%3Csup%3E%C2%AE%3C-sup%3E-MagBeads-Streptavidin/p/0000000a0002a23a00010023_de"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
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<li>Company: Biotium</li> | <li>Company: Biotium</li> | ||
<li>REF: 41003</li> | <li>REF: 41003</li> | ||
| − | + | <a href="https://biotium.com/product/gelredtm-nucleic-acid-gel-stain-10000x-in-water/"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 335: | Line 335: | ||
<li>Company: Affymetrix eBioscience</li> | <li>Company: Affymetrix eBioscience</li> | ||
<li>REF: BMS147</li> | <li>REF: BMS147</li> | ||
| − | + | <a href="http://www.ebioscience.com/human-annexin-v-antibody-biotin-vaa-33.htm"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 350: | Line 350: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: B7025S</li> | <li>REF: B7025S</li> | ||
| − | + | <a href="https://www.neb.com/products/b7025-gel-loading-dye-purple-6x-no-sds"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 365: | Line 365: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: M0202S</li> | <li>REF: M0202S</li> | ||
| − | + | <a href="https://www.neb.com/products/m0202-t4-dna-ligase"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 380: | Line 380: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: M0140S</li> | <li>REF: M0140S</li> | ||
| − | + | <a href="https://www.neb.com/products/r0140-psti"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 395: | Line 395: | ||
<li>Company: New England BioLabs</li> | <li>Company: New England BioLabs</li> | ||
<li>REF: B0208S</li> | <li>REF: B0208S</li> | ||
| − | + | <a href="https://www.neb.com/products/b0208-taq-dna-ligase-reaction-buffer"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 410: | Line 410: | ||
<li>Company: </li> | <li>Company: </li> | ||
<li>REF: </li> | <li>REF: </li> | ||
| − | + | <a href=""><p>for further information click here</p></a> | |
</ul> | </ul> | ||
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<li>Company: </li> | <li>Company: </li> | ||
<li>REF: </li> | <li>REF: </li> | ||
| − | + | <a href=""><p>for further information click here</p></a> | |
</ul> | </ul> | ||
| Line 438: | Line 438: | ||
<li>Company: Sigma-Aldrich</li> | <li>Company: Sigma-Aldrich</li> | ||
<li>REF: 02860-2.5L</li> | <li>REF: 02860-2.5L</li> | ||
| − | + | <a href="http://www.sigmaaldrich.com/catalog/product/fluka/02860?lang=de®ion=CH"><p>for further information click here</p></a> | |
</ul> | </ul> | ||
Revision as of 15:27, 8 September 2015
- Project
- Modeling
- Lab
- Human
Practices - Parts
- About Us
Materials
Buffers and media
TFP1
- 100 mM RbCl
- 50 mM MnCl2
- 30 mM potassium acetate
- 10 mM CaCl2
- 15%glycerol
TFP2
- 100 mM MOPS
- 50 mM RbCl
- 75 mM CaCl2
- 15%glycerol
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.
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 )
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
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.
Chemicals and Kits
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
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
Streptavidin Beads
- Product: Roti®-MagBeads Streptavidin
- Company: Carl Roth
- REF:
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
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).
Bacterial strains and Mammalian Cell Lines
TOP10
The bacterial strain we used as a base for all our transformations was TOP10.
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).
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).
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.
SK-BR-3
HER2-positive breast cancer cell line. We used this cell line to validate the chimeric receptors of the Stockholm iGEM team.
Machines
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
Fluorescence Scanner for Western Blots
- ODYSSEY® CLx Infrared Imaging System
for further information click here
Emulsifier
- EmulsiFlex-C3 from AVESTIN, Inc. and AVESTIN Europe GmbH/li>
for further information click here