Protocols

Protocols organised by category

Biochemistry

• GUS assay (β-Glucuronidase assay)

Microbiology

• Characterisation of Actinomycetes

• General Growth and Maintenance of Streptomyces sp.

• Isolating Actinomycetes from the environment

Microscopy

• Staining of Streptomyces spp. Cell Wall and Nucleic Acids

Mutagenesis

• CRISPR/Cas9-mediated genome editing

• Chemical mutagenesis

• Depletion strains

• Lambda-red mediated recombination (PCR-targeting system a.k.a. "Redirect")

• Lambda-red mediated recombination using ssDNA

• Meganuclease I-SceI based system for gene deletions

• "Quikchange" site-directed mutagenesis

• Random mutagenesis using a mutator strain

• Random mutagenesis using error-prone PCR

• Suicide vectors

• Transposons

• UV mutagenesis

Natural Products

• Cloning, Transformation and Expression of Gene Clusters

• Metabolite extractions

Next Generation Sequencing

• ChIP Sequencing
• ChIP-exo sequencing

https://www.nature.com/articles/s41467-018-05265-7/

• RNA Sequencing
• Differential RNA sequencing

Nucleic Acid Methods

• Codon Optimising Sequences for Heterologous Expression in Streptomyces

• High GC PCR

• RNA extraction from Actinobacteria

• Salting Out Genomic DNA Extraction Method

• Southern Blotting

Protein Methods

• Isolating extracellular protein from Streptomyces

Alphabetical list of protocols

Characterisation of Actinomycetes

Cloning, Transformation and Expression of Gene Clusters

CRISPR/Cas9-mediated genome editing

Chemical mutagenesis

ChIP Seq

Codon Optimising Sequences for Heterologous Expression in Streptomyces

Depletion strains

General Growth and Maintenance of Streptomyces sp.

GUS assay (β-Glucuronidase assay)

High GC PCR

Isolating Actinomycetes from the environment

Isolating extracellular protein from Streptomyces

Lambda-red mediated recombination (PCR-targeting system a.k.a. "Redirect")

Lambda-red mediated recombination using ssDNA

Meganuclease I-SceI based system for gene deletions

Metabolite extractions

"Quikchange" site-directed mutagenesis

Random mutagenesis using a mutator strain

Random mutagenesis using error-prone PCR

RNA extraction from Actinobacteria

Salting Out Genomic DNA Extraction Method

Southern Blotting

Staining of Streptomyces spp. Cell Wall and Nucleic Acids

Suicide vectors

Transposons

UV mutagenesis