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</div> | </div> | ||
− | <div class="contentbox"> | + | <div class="contentbox"> |
<div class="content"> | <div class="content"> | ||
<h3> Protocols </h3> | <h3> Protocols </h3> | ||
+ | <div class="article"> | ||
+ | This page lists all regular methods used during our project. Special methods, which are not listed here, are described or linked on the corresponding result pages. | ||
+ | </div> | ||
<div class="contentline"> | <div class="contentline"> | ||
<div class="half left"> | <div class="half left"> | ||
<ul style="list-style-type: none"> | <ul style="list-style-type: none"> | ||
<li><a href="https://static.igem.org/mediawiki/2017/8/85/T--Bielefeld-CeBiTec--protocol_50xTAE.pdf" target="blank">50xTAE Stock Solution</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/8/85/T--Bielefeld-CeBiTec--protocol_50xTAE.pdf" target="blank">50xTAE Stock Solution</a></li> | ||
− | <li><a href="https://static.igem.org/mediawiki/2017/ | + | <li><a href="https://static.igem.org/mediawiki/2017/6/61/T--Bielefeld-CeBiTec--CMZ-Acetone.pdf" target="blank">Acetone Precipitation</a></li> |
− | <li><a href="https://static.igem.org/mediawiki/2017/ | + | <li><a href="https://static.igem.org/mediawiki/2017/a/ae/T--Bielefeld-CeBiTec--CMZ-Biotin.pdf" target="blank">Biotinylation of glass slides</a></li> |
<li><a href="https://static.igem.org/mediawiki/2017/3/3a/T--Bielefeld-CeBiTec--protocol_blunt-end-ligation.pdf" target="blank">Blunt end ligation</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/3/3a/T--Bielefeld-CeBiTec--protocol_blunt-end-ligation.pdf" target="blank">Blunt end ligation</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/6/6f/T--Bielefeld-CeBiTec--RotiNanoquant.pdf">Roti_Nanoquant (for protein concentration estimation)</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/9/97/T--Bielefeld-CeBiTec--protocol_cloning_strategy_re.pdf" target="blank">Cloning Strategy with Restriction Enzymes</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/c/cc/T--Bielefeld-CeBiTec--protocol_colonyPCR_GoTaq_G2.pdf" target="blank">Colony PCR GoTaq® G2 DNA Polymerase</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/c/cc/T--Bielefeld-CeBiTec--protocol_colonyPCR_GoTaq_G2.pdf" target="blank">Colony PCR GoTaq® G2 DNA Polymerase</a></li> | ||
− | <li><a href="https://static.igem.org/mediawiki/2017/ | + | <li><a href="https://static.igem.org/mediawiki/2017/2/25/T--Bielefeld-CeBiTec--CMZ-FastcellSDS.pdf" target="blank">Fast cell lysis sample buffer</a></li> |
− | + | ||
<li><a href="https://static.igem.org/mediawiki/2017/e/e0/T--Bielefeld-CeBiTec--protocol_fast_cell_lysis_SDS-Page.pdf" target="blank">Fast cell lysis for SDS-PAGE</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/e/e0/T--Bielefeld-CeBiTec--protocol_fast_cell_lysis_SDS-Page.pdf" target="blank">Fast cell lysis for SDS-PAGE</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/3/39/T--Bielefeld-CeBiTec--protocol_GeneJET_DNA_Purification_Thermo.pdf.pdf" target="blank">GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific)</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/3/39/T--Bielefeld-CeBiTec--protocol_GeneJET_DNA_Purification_Thermo.pdf.pdf" target="blank">GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific)</a></li> | ||
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<li><a href="https://static.igem.org/mediawiki/2017/4/42/T--Bielefeld-CeBiTec--protocol_generating_chemocompetent_cells.pdf" target="blank">Generating chemocompetent cells</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/4/42/T--Bielefeld-CeBiTec--protocol_generating_chemocompetent_cells.pdf" target="blank">Generating chemocompetent cells</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/6/6c/T--Bielefeld-CeBiTec--protocol_generating_electrocompetent_cells.pdf" target="blank">Generating electrocompetent cells</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/6/6c/T--Bielefeld-CeBiTec--protocol_generating_electrocompetent_cells.pdf" target="blank">Generating electrocompetent cells</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/3/37/T--Bielefeld-CeBiTec--protocol_genome_editing_codA.pdf">Genome editing via the CRISP-Cas9 system</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/5/55/T--Bielefeld-CeBiTec--protocol_gibson_assembly.pdf" target="blank">Gibson Assembly</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/5/55/T--Bielefeld-CeBiTec--protocol_gibson_assembly.pdf" target="blank">Gibson Assembly</a></li> | ||
− | + | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/3/3f/T--Bielefeld-CeBiTec--CMZ-Membrane.pdf" target="blank">Isolation of the Inner Membrane Fraction</a></li> | ||
+ | |||
+ | |||
<li><a href="https://static.igem.org/mediawiki/2017/7/75/T--Bielefeld-CeBiTec--protocol_innuPREP_Gel_analytikJena.pdf" target="blank">innuPREP Gel Extraktion Kit (Analytik Jena)</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/7/75/T--Bielefeld-CeBiTec--protocol_innuPREP_Gel_analytikJena.pdf" target="blank">innuPREP Gel Extraktion Kit (Analytik Jena)</a></li> | ||
− | <li><a href="https://static.igem.org/mediawiki/2017/a/a7/T--Bielefeld-CeBiTec--protocol_LB.pdf" target="blank"> | + | <li><a href="https://static.igem.org/mediawiki/2017/8/8c/T--Bielefeld-CeBiTec--YKE_lycopene_protocol.pdf" target="blank">Lycopene extraction</a></li> |
+ | <li><a href="https://static.igem.org/mediawiki/2017/a/a7/T--Bielefeld-CeBiTec--protocol_LB.pdf" target="blank">Lysogeny Broth (LB)</a></li> | ||
+ | |||
+ | <li><a href="https://static.igem.org/mediawiki/2017/a/a4/T--Bielefeld-CeBiTec--CMZ-Maldi.pdf" target="blank">Malditof sample preparation</a></li> | ||
+ | |||
+ | <li><a href="https://static.igem.org/mediawiki/2017/8/8a/T--Bielefeld-CeBiTec--protocol_Monarch_Miniprep_kit_NEB.pdf" target="blank">Monarch® Plasmid Miniprep Kit (NEB)</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/0/0e/T--Bielefeld-CeBiTec--protocol_Monarch_PCR_and_DNA_clean_up_kit_NEB.pdf" target="blank">Monarch® PCR & DNA Cleanup Kit (NEB)</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/7/7d/T--Bielefeld-CeBiTec--protocol_MOPS.pdf" target="blank">MOPS Minimal Medium</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
<div class="half right"> | <div class="half right"> | ||
<ul style="list-style-type: none"> | <ul style="list-style-type: none"> | ||
− | |||
− | |||
− | |||
<li><a href="https://static.igem.org/mediawiki/2017/e/e6/T--Bielefeld-CeBiTec--protocol_native_PAGE.pdf" target="blank">Native DNA PAGE</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/e/e6/T--Bielefeld-CeBiTec--protocol_native_PAGE.pdf" target="blank">Native DNA PAGE</a></li> | ||
− | <li><a href="https://static.igem.org/mediawiki/2017/6/ | + | <li><a href="https://static.igem.org/mediawiki/2017/6/62/T--Bielefeld-CeBiTec--CMZ-PBJRbuffer.pdf" target="blank">PBJR buffer</a></li> |
− | <li><a href="https://static.igem.org/mediawiki/2017/ | + | <li><a href="https://static.igem.org/mediawiki/2017/2/2b/T--Bielefeld-CeBiTec--ProtocolPhusion.pdf" target="blank">PCR with Phusion High Fidelity Master Mix with HC enhancer</a></li> |
− | <li><a href="https://static.igem.org/mediawiki/2017/ | + | <li><a href="https://static.igem.org/mediawiki/2017/1/16/T--Bielefeld-CeBiTec--CMZ-PCRQ5.pdf" target="blank">PCR with Q5 High-Fidelity Polymerase</a></li> |
+ | <li><a href="https://static.igem.org/mediawiki/2017/e/ec/T--Bielefeld-CeBiTec--CMZ-GoldenGate.pdf" target="blank">Plasmid Assembly Protocol with Golden Gate Assembly</a></li> | ||
+ | |||
+ | <li><a href="https://static.igem.org/mediawiki/2017/d/d7/T--Bielefeld-CeBiTec--CMZ-Mikroskopie.pdf" target="blank">Protocol for GFP immunofluorescence (whole Bacteria)</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/0/0e/T--Bielefeld-CeBiTec--ImpactProtocol.pdf">Protein Purification Impact-CN Protocol</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/9/96/T--Bielefeld-CeBiTec--protocol_PCR_PureYield_PlasmidMiniprep_Promega.pdf" target="blank">PureYield<sup>TM</sup> Plasmid Miniprep System (Promega)</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/9/96/T--Bielefeld-CeBiTec--protocol_PCR_PureYield_PlasmidMiniprep_Promega.pdf" target="blank">PureYield<sup>TM</sup> Plasmid Miniprep System (Promega)</a></li> | ||
− | <li><a href="https://static.igem.org/mediawiki/2017/ | + | <li><a href="https://static.igem.org/mediawiki/2017/0/0d/T--Bielefeld-CeBiTec--protocol_SDS-PAGE.pdf" target="blank">SDS-PAGE</a></li> |
<li><a href="https://static.igem.org/mediawiki/2017/a/a2/T--Bielefeld-CeBiTec--protocol_SDS-PAGE_running_buffer.pdf" target="blank">SDS-PAGE running buffer</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/a/a2/T--Bielefeld-CeBiTec--protocol_SDS-PAGE_running_buffer.pdf" target="blank">SDS-PAGE running buffer</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/1/19/T--Bielefeld-CeBiTec--protocol_SiO2_column.pdf" target="blank">SiO2 Column Chromatography with Gradient</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/f/fe/T--Bielefeld-CeBiTec--protocol_SOC.pdf" target="blank">SOC-medium</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/f/fe/T--Bielefeld-CeBiTec--protocol_SOC.pdf" target="blank">SOC-medium</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/d/d1/T--Bielefeld-CeBiTec--protocol_ssDNA_annealing.pdf" target="blank">ssDNA Annealing</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/d/d1/T--Bielefeld-CeBiTec--protocol_ssDNA_annealing.pdf" target="blank">ssDNA Annealing</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/8/84/T--Bielefeld-CeBiTec--protocol_Standard_Biobrick.pdf" target="blank">Standard BioBrick Assembly</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/8/84/T--Bielefeld-CeBiTec--protocol_Standard_Biobrick.pdf" target="blank">Standard BioBrick Assembly</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/a/a6/T--Bielefeld-CeBiTec--protocol_CBT-asparagine.pdf" target="blank">Synthesis of CBT-asparagine</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/d/d4/T--Bielefeld-CeBiTec--protocol_TAE_buffer.pdf" target="blank">TAE buffer</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/d/d4/T--Bielefeld-CeBiTec--protocol_TAE_buffer.pdf" target="blank">TAE buffer</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/a/a2/T--Bielefeld-CeBiTec--protocol_TLC.pdf" target="blank">TLC analysis</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/a/ab/T--Bielefeld-CeBiTec--protocol_electrotrafo.pdf" target="blank">Transformation via electroporation</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/a/a0/T--Bielefeld-CeBiTec--protocol_heatshock.pdf" target="blank">Transformation via heat shock</a></li> | ||
+ | <li><a href="https://static.igem.org/mediawiki/2017/2/23/T--Bielefeld-CeBiTec--protocol_Restriction_Digest.pdf" target="blank">"Typical" Restriction Digest</a></li> | ||
+ | |||
+ | <li><a href="https://static.igem.org/mediawiki/2017/8/88/T--Bielefeld-CeBiTec--CMZ-Westernblot.pdf" target="blank">Western Blot</a></li> | ||
+ | |||
+ | <li><a href="https://static.igem.org/mediawiki/2017/2/2e/T--Bielefeld-CeBiTec--protocol_PCR_Gel_Cleanup_wizzard.pdf" target="blank">Wizard® SV Gel and PCR Clean-Up System (Promega)</a></li> | ||
<li><a href="https://static.igem.org/mediawiki/2017/1/13/T--Bielefeld-CeBiTec--protocol_ZymoPURETM_PlasmidMiniprep_Kit.pdf" target="blank">ZymoPURE<sup>TM</sup> Plasmid Miniprep Kit (Zymo Research)</a></li> | <li><a href="https://static.igem.org/mediawiki/2017/1/13/T--Bielefeld-CeBiTec--protocol_ZymoPURETM_PlasmidMiniprep_Kit.pdf" target="blank">ZymoPURE<sup>TM</sup> Plasmid Miniprep Kit (Zymo Research)</a></li> | ||
− | |||
</ul> | </ul> | ||
</div> | </div> | ||
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<tr> | <tr> | ||
<td><a href="https://www.thermofisher.com/order/catalog/product/ER0301"><i>Eco</i>RV</a></td> | <td><a href="https://www.thermofisher.com/order/catalog/product/ER0301"><i>Eco</i>RV</a></td> | ||
− | <td> | + | <td>Thermo Fisher Scientific</td> |
</tr> | </tr> | ||
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</thead> | </thead> | ||
<tbody> | <tbody> | ||
− | + | <tr> | |
− | + | <td><a target="_blank"href="https://www.highqu.com/allintmhifi-dna-polymerase-2-u-l.html">Allin HiFi DNA polymerase</a></td> | |
+ | <td>highQu</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><a target="_blank"href=" http://biolabmix.ru/en/products/klassicheskaja_pcr/biomaster_hs-taq_pcr-color__2_/"> BioMaster-HS Taq </a></td> | ||
+ | <td>Biolabmix</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><a target="_blank"href=" https://www.sbd.ee/EN/products/pcr/firepol "> FirePol DNA polymerase </a></td> | ||
+ | <td>Solis Biodyne</td> | ||
+ | </tr> | ||
<tr> | <tr> | ||
<td><a href="https://www.promega.de/products/pcr/endpoint-pcr/gotaq-dna-polymerase/?catNum=M3001">Go Taq DNA polymerase</a></td> | <td><a href="https://www.promega.de/products/pcr/endpoint-pcr/gotaq-dna-polymerase/?catNum=M3001">Go Taq DNA polymerase</a></td> | ||
<td>Promega</td> | <td>Promega</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
+ | <td><a target="_blank"href="https://www.analytik-jena.de/fileadmin/content/import/imported_dam/br_Product_Guide_Reagents_Plastic_WEB_geschuetzt.pdf">innuDRY DNA polymerase </a></td> | ||
+ | <td>Analytik Jena</td> | ||
+ | </tr> | ||
+ | |||
<tr> | <tr> | ||
Line 166: | Line 208: | ||
<td>Thermo Fisher Scientific</td> | <td>Thermo Fisher Scientific</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
+ | <td><a href="https://www.neb.com/products/m0530-phusion-high-fidelity-dna-polymerase#Product%20Information">Phusion High-Fidelity DNA polymerase </a></td> | ||
+ | <td>NEB</td> | ||
+ | </tr> | ||
+ | |||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<tr> | <tr> | ||
<td><a href="https://www.neb.com/products/m0491-q5-high-fidelity-dna-polymerase">Q5 High-Fidelity DNA polymerase </a></td> | <td><a href="https://www.neb.com/products/m0491-q5-high-fidelity-dna-polymerase">Q5 High-Fidelity DNA polymerase </a></td> | ||
<td>NEB</td> | <td>NEB</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
+ | <td><a target="_blank"href="http://www.clontech.com/DE/Products/PCR/High_Yield_PCR/Titanium_Taq_DNA_Polymerase?sitex=10024:22372:US">Titanium Taq</a></td></td> | ||
+ | <td>Clonetech</td> | ||
+ | </tr> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <tr> | ||
+ | |||
+ | |||
− | |||
− | |||
− | |||
− | |||
</tbody> | </tbody> | ||
</table> | </table> |
Latest revision as of 03:29, 2 November 2017
Methods
Protocols
This page lists all regular methods used during our project. Special methods, which are not listed here, are described or linked on the corresponding result pages.
- 50xTAE Stock Solution
- Acetone Precipitation
- Biotinylation of glass slides
- Blunt end ligation
- Roti_Nanoquant (for protein concentration estimation)
- Cloning Strategy with Restriction Enzymes
- Colony PCR GoTaq® G2 DNA Polymerase
- Fast cell lysis sample buffer
- Fast cell lysis for SDS-PAGE
- GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific)
- GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific)
- Generating chemocompetent cells
- Generating electrocompetent cells
- Genome editing via the CRISP-Cas9 system
- Gibson Assembly
- Isolation of the Inner Membrane Fraction
- innuPREP Gel Extraktion Kit (Analytik Jena)
- Lycopene extraction
- Lysogeny Broth (LB)
- Malditof sample preparation
- Monarch® Plasmid Miniprep Kit (NEB)
- Monarch® PCR & DNA Cleanup Kit (NEB)
- MOPS Minimal Medium
- Native DNA PAGE
- PBJR buffer
- PCR with Phusion High Fidelity Master Mix with HC enhancer
- PCR with Q5 High-Fidelity Polymerase
- Plasmid Assembly Protocol with Golden Gate Assembly
- Protocol for GFP immunofluorescence (whole Bacteria)
- Protein Purification Impact-CN Protocol
- PureYieldTM Plasmid Miniprep System (Promega)
- SDS-PAGE
- SDS-PAGE running buffer
- SiO2 Column Chromatography with Gradient
- SOC-medium
- ssDNA Annealing
- Standard BioBrick Assembly
- Synthesis of CBT-asparagine
- TAE buffer
- TLC analysis
- Transformation via electroporation
- Transformation via heat shock
- "Typical" Restriction Digest
- Western Blot
- Wizard® SV Gel and PCR Clean-Up System (Promega)
- ZymoPURETM Plasmid Miniprep Kit (Zymo Research)
Enzymes
Digestion enzymes
Enzyme | Producer |
---|---|
AgeI-HF | NEB |
BsaI | NEB |
DpnI | NEB |
DpnI | Jena Bioscience |
EciI | NEB |
EcoRV | Thermo Fisher Scientific |
MnlI | NEB |
NotI | Jena Bioscience |
PstI | NEB |
PstI | Thermo Fisher Scientific |
PstI-HF | NEB |
SapI | NEB |
SpeI | NEB |
XbaI | NEB |
XbaI | Jena Bioscience |
Polymerases
Enzyme | Producer |
---|---|
Allin HiFi DNA polymerase | highQu |
BioMaster-HS Taq | Biolabmix |
FirePol DNA polymerase | Solis Biodyne |
Go Taq DNA polymerase | Promega |
innuDRY DNA polymerase | Analytik Jena |
Klenow Fragment | Thermo Fisher Scientific |
Phusion High-Fidelity DNA polymerase | NEB |
Q5 High-Fidelity DNA polymerase | NEB |
Titanium Taq | Clonetech |
Other Enzymes
Enzyme | Producer |
---|---|
Shrimp Alkaline Phosphatase | Jena Bioscience |
T4 DNA Ligase | NEB |
T4 DNA Ligase | Thermo Fisher Scientific |
T4 Polynucleotide Kinase | NEB |