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Getting settled in the Swartz Lab

Contents 1 Protocols 1.1 Lab Ordering 1.2 Cloning 1.3 Cell Free Protein Synthesis 1.4 Protein Purification 1.5 Assays 1.6 Miscellaneous 1.7 Solutions

Protocols

Lab Ordering

• USE THIS FORM

Cloning

• Swartz Lab Molecular Biology Handbook
• Restriction Enzyme Library
• Gibson_Assembly

DNA Design

• Primer Design

Gene Synthesis

• PCR
• Gene Synthesis
• Restriction Enzyme Digestion/Ligations
• Quikchange
• Gel Purification/PCR Cleanup
• MaxiPrep Column Regeneration
• Genomic DNA Extraction

Expression Templates

Plasmids

• OWW Plasmid Miniprep or | Swartz Lab Version
• Plasmid Maxiprep

Organisms

• E.coli Transformation
• Other Cloning Protocols
• Creating an E. coli glycerol stock

Cell Free Protein Synthesis

• Preparing linear DNA template for CFPS
• 1L Extract Prep
• Cell Free Protein Synthesis
• How to do an Autoradiogram
• Western Blotting
• High Density Fermentation
• Orthogonal tDNA template prep
• Running a protein gel

• • S30_buffer

Protein Purification

• His-Tag Purification
• Ribosome purification

Assays

• Nanodrop
• Using the spec for DNA Quantification
• Sample prep for MS
• Spin assay for ribsome binding
• HPLC
• Ellman's Reagent Test
• Cytochrome C
• Methyl Viologen
• H2 Production (GC)

Miscellaneous

• Anaerobic Protein Production in E.coli
• Cell culture for A20 B-cells

Solutions

• Buffer Recipes