Difference between revisions of "Team:Bielefeld-CeBiTec/Notebook/Methods"
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<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/2017/3/37/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> | ||
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| + | <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> | ||
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<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/2017/8/8c/T--Bielefeld-CeBiTec--YKE_lycopene_protocol.pdf" target="blank">Lycopene extraction</a></li> | <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/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> | ||
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<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/2017/8/8a/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/2017/0/0e/T--Bielefeld-CeBiTec--protocol_Monarch_PCR_and_DNA_clean_up_kit_NEB.pdf" target="blank">Monarch® PCR & DNA Cleanup Kit (NEB)</a></li> | ||
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<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/2017/1/16/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/e/ec/T--Bielefeld-CeBiTec--CMZ-GoldenGate.pdf" target="blank">Plasmid Assembly Protocol with Golden Gate Assembly</a></li> | ||
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| + | <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/2017/0/0e/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> | ||
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<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/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/2017/2/23/T--Bielefeld-CeBiTec--protocol_Restriction_Digest.pdf" target="blank">"Typical" Restriction Digest</a></li> | ||
| − | <li><a href="https://static.igem.org/mediawiki/2017/2/2e/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/2017/8/88/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> | ||
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 |
