Protocols

Growth of Salmonella typhimurium or Escherichia coli.

Preparation of antibiotics

Resuccitation of -80°C freezer culture

Over-night starter culture

Culture for experiments

RNA isolation, hot phenol

Solutions

RNeasy RNA isolation

RNeasy Mini RNA prep protocol

DNase Step

RNeasy Mini RNA prep protocol continued

Solutions

Control of RNA

The amounts and the quality of the RNA is checked in the spectrophotometer and on a denaturing RNA-gel.

Measurements in spectrophotometer

RNA-gel

Picture of RNA-gel

Solutions

Production and purification of labeled cDNA

Reverse transcription with Superscript II and labelling with Cy-dyes

Purification on Microspin column

Solutions

Hybridisation

Preparation of slide for Hybridisation

Hybridisation of cDNA to slide

Stringency washes

Solutions

Scanning

Scanning of slide on arrayWoRxe scanner

Genomic DNA labelling for microarray quality control

Reverse PCR for mRNA confirmation

Primer Extension to confirm results or examine expression of single mRNAs

PCR protocol

Making competent cells of Escherichia coli or Salmonella typhimurium, and transformation of these by electroporation

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