SOPs

Sample preparation procedure for metabolic analysis on T. b. brucei 427 using LC/MS

For the study of mRNA decay rates, transcription was inhibited with ActinomycinD, and RNA splicing with Sinefungin, at different time points, in the Matthews lab. rRNA depleted RNA was extracted from each of the samples in the Clayton lab, and sent for deep sequencing at the BioQuant facility in Heidelberg

Sample preparation procedure for metabolic footprint analysis on T. b. brucei 427 using LC/MS

Biomass sampling - SOP for sampling of biomass

This is a well-established, classical genetic method of constructing chromosomal monolysogenic fusions to a promoterless lacZ gene.

Method extraction of intracellular metabolites in Lactococcus lactis

The reverse transcriptase synthesizes DNA, which complements the mRNA template (complementary DNA, cDNA). Cy3/Cy5-dCTP are incorporated into cDNA during Reverse transcription. The obtained Cy3/Cy5 cDNA are then competitively hybridised onto Agilent microarray slide and subsequently scanned.

SOP for shake flask cultivation of B.Subtilis in Bacell-Sysmo

ClosTron mutants should always be subjected to Southern blot analysis to ensure that only one intron insertion has occurred.

SOPs related with medium composition, fermentation and stocking of S. solfataricus.

Standard procedure regarding intracellular metabolome analysis using GC-MS.

Protocol for RNA isolation, cDNA synthesis and labeling and hybridization and cleanup of Sulfolobus solfataricus microarray

Standard operating procedures regarding iTraq based proteomics.

This SOP describes the SUMO procedure for determining B-galactosidase activities.

This document describes the chemostat conditions to run comparable experiments in different bioreactors in different labs.

This file describes how to isolate mRNA from E. coli using the kit from Epicentre, for gene expression analysis via RT-PCR

This SOP describes the preparation of E. coli RNA

This protocol describes the transcriptional profiling of E. coli cultures using microarrays. The protocol utilises RNA isolated as described in another SOP (SUMO RNA isolation from E. coli) and with hybridisation to Ocimum Ocichip E. coli K-12 microarrays.