Team:Munich/Notebook

Notebook

Cell Extract Optimization

• 09:00April
• 09:30April
• 10:00May
• 10:30May
• 11:00June
• 11:30June
• 12:00July
• 12:30July
• 13:00August
• 13:30August
• 14:00September
• 14:30September
• 15:00October
• 15:30October

• Cultivation
  • Screening Additional Parameters
  • Evaluating E. Coli Rosetta 2 Growth
  • Screening at Different OD600s
  • Screening Knockout Strains for Cell Extract Quality
• Cell Lysis
  • Screening Lysis Methods
  • Testing different plasmid Preparation methods
  • Screening Sonication Parameters
  • Final Settings
  • Biological Triplicates
• Lyophilisation And Upscaling
  • Exploring the Possibilities of Upscaling
  • Proof of Principle: Cell Extract Lyophilsation
• Quality Control
  • Collaboration with Mexico
  • Cloning our QC Plasmid

Close

Cell Extract Engineering

• 09:00April
• 09:30April
• 10:00May
• 10:30May
• 11:00June
• 11:30June
• 12:00July
• 12:30July
• 13:00August
• 13:30August
• 14:00September
• 14:30September
• 15:00October
• 15:30October

• Chi6 Inhibitor for RecBCD
  • Creating a Biobrick for Chi6
  • Versions of the Chi6 Inhibitor
  • RecBCD Inhibition In-Vitro
  • Creating a Biobrick for Chi6
  • RecBCD Inhibition in Cell Extract
• Genome Engineering
  • Preparing pRED/ET Engineering
  • Creating a Selection Cassette for Deletions
  • pRED/ET Engineering in E. Coli Rosetta
  • Preparing pRED/ET Engineering
  • Creating a Selection Cassette for Deletions
  • pRED/ET Genome Deletions in E.Coli MG1655
  • pRED/ET Engineering in E. Coli Rosetta
• RecBCD Engineering
  • Genomic DNA Extraction
  • Cloning RecBCD WT + His
  • Engineering TEV-sites into RecBCD
  • Cloning RecBCD WT
  • Engineering TEV-sites into RecBCD
  • Creating Biobricks for RecB, C, D
  • Inhibiting RecBCD using dCas9
  • Evaluating the Growth of E.Coli Knockouts
• Cas13a
  • cas13a Based Selection System for MAGE
  • Backup Gblock sgRNA against EndoA 6-7

Close

Phage Preparation

• 09:00April
• 09:30April
• 10:00May
• 10:30May
• 11:00June
• 11:30June
• 12:00July
• 12:30July
• 13:00August
• 13:30August
• 14:00September
• 14:30September
• 15:00October
• 15:30October

• Phage-DNA Production
  • DNA Purification through phenol chloroform precipitation (T7 and MS2 Phage)
  • DNA isolation with Phage DNA isolation Kit and improved Phenol chloroform precipitations (T7, T4 and 3S Phage)
  • Preparing T4, 3S, NES and FFP Phages for sequencing
• Phage Assembly
  • T7 and MS2 Phage Assembly in commercial and home-made cell extract
  • Phage Assemblies and Burst Size Analysis
  • Phage Assembly in home-made cell extract
• Modular Phage Composition
  • QPCR
  • gRNA Generation
  • Cloning of N-HOC-His-C
  • Phage Genome Engineering
  • Creating BioBricks for Phage Proteins
  • His-HOC-YFP Purification and Assembly
• Other
  • OraColi
  • Quality Control by qPCR
  • Encapsulation
  • NanoPore Sequencing

Close

Primer List

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