Team:Freiburg/Protocols

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Cloning

• Blunt End Ligation
• Competent Cells
• Cryo Stocks
• gBlock Procedure
• Agarose Gels
• Antibiotics
• Colony PCR
• Competent Cells
• Fusion PCR
• Gel Extraction
• Gibson Assembly
• Isopropanol Precipitation
• Ligation
• Miniprep
• PCR
• Restriction Digest
• Transformation

Cell-free Expression

• Cell-free Expression on PDMS Flow Cell
• Preparation of Terrific Broth
• Cell-free Expression general
• Preparation of an E.Coli lysate for Cell-free Expression
• Luciferase Assay
• Measurement of GFP Expression

Surface Chemistry

• Plasma Activation
• GOPTS Surface
• APTES-PDITC Surface
• Ni-NTA Surface
• HALO Surface (Silane)
• Glass Slide Preparation (full)
• iRIF Slide Preparation (full)
• Spotting
• Moulding Flowcells
• PDITC Surface on PDMS

Protein Purification & Analysis

• AmidoBlack
• Ni-NTA Bead Regeneration
• Buffers for Analysis
• Cell Lysis
• Desalting of Protein Solutions (Columns)
• DotBlot
• Freezing and Thawing Cells
• Microwave Destaining
• Overexpression
• Periplasmic Preparation
• Purification
• SDS-Page
• Blood Serum Preparation
• Transfection
• Western Blot

iRIf

• iRIf Measurement
• Protein Stamping
• iRIf Slide Cleaning

Interlab Study

• InterLab Protocol & Worksheet