The Coleman LabThe Coleman Lab

Lab Protocols

General lab safety and aseptic technique

Making agar plates

MSM minimal medium

Growing E.coli 

Making, running, staining agarose gels

Preparation of chemically-competent E.coli

Heat shock transformation of chemically-competent E.coli

Preparation of electrocompetent cells of Pseudomonas or E.coli, and electroporation

Preparation of electrocompetent cells of Mycobacterium, and electroporation

Genomic DNA extraction with CTAB

Genomic DNA extraction with bead beater

Resting cell experiments with bacteria

Plasmid miniprep - 5 ml - spin column

Plasmid midiprep - 50 ml - spin column

Plasmid midiprep - 50 ml - phenol-chloroform

Polymerase chain reaction (PCR) and primer design

Purification of PCRs or digests via spin column

Purification of PCRs or digests from gel by spin column

Quantitation and digestion of plasmid DNA 

Golden Gate Assembly of DNA

Cloning into plasmids - digestion, ligation, troubleshooting

Screening recombinant clones (patch, PCR, digest)

Validation of recombinant plasmids by Sanger Sequencing

Protein analysis by SDS-PAGE

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