Protocols

From ActinoBase

Protocols

Protocols organised by category

Bacteriophage

Biochemistry

Microbiology

For a comprehensive guide to actinomycete characterisation see the Compendium of Actinobacteria from Prof. Joachim Wink at the German Collection of Microorganisms & Cell Cultures (DSMZ). This resouce provides protocols for characterising a wide range of features for new isolates, but may also be applicable more broadly for example for phenotyping experiments. The tests include colony morphology, melanin production, salt tolerance, carbon utlilisation and much more. To download the guide please see the link below.

DSMZ Compendium of Actinobacteria

Microscopy

Mutagenesis

Natural Products

Next Generation Sequencing

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

Nucleic Acid Methods

Protein Methods

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

Conjugation using ET12567/pUZ8002

Depletion strains

Streptomyces Exploration

General Growth and Maintenance of Streptomyces sp.

GUS assay (β-Glucuronidase assay)

High GC PCR

High titre preparation of phage

Isolating Actinomycetes from the environment

Isolating extracellular protein from Streptomyces

Isolation of phages from environmental samples

Isolation of phages from single plaques

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

Protoplasts Formation, Regeneration and Transformation

"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

Streptomyces Bacteriophage plaque assay

Suicide vectors

Transposons

UV mutagenesis