SOPs
What is a SOP?Isolation of total RNA from Bacillus Subtilis using phenol-chloroform extraction method by maintaining cryogenec conditions initailly to prevent RNA degradation. Quality of the obtained RNA is then tested with Agilent Bioanalyser before proceeding for gene expression analysis.
Creator: Praveen kumar Sappa
Submitter: Praveen kumar Sappa
An overview of creating MAGE-TAB compliant spreadsheets for transcriptomics data in SysMO SEEK
Creator: Katy Wolstencroft
Submitter: Katy Wolstencroft
The procedure describes the preparation of fluorescent DNA probes from human mRNA or total RNA.
Creator: Olga Krebs
Submitter: Olga Krebs
This is a protocol for determining the activity of the T. brucei Trypanothione synthetase under in vivo-like conditions.
Creators: Alejandro Leroux, Luise Krauth-Siegel
Submitter: Alejandro Leroux
This method describes the preparation of the in vivo-like buffer for the measurement of bloodstream T. brucei recombinant enzymes under pseudo-physiological conditions.
Creators: Alejandro Leroux, Luise Krauth-Siegel
Submitter: Alejandro Leroux
Preparation of cell free extracts of the recombinant E. coli strains expressing the gluconeogenic S. solfataricus enzymes.
Creators: Jacky Snoep, Theresa Kouril
Submitter: Jacky Snoep
Cloning and heterologous expression of gluconeogenic enzymes from S. solfataricus in E. coli
Creators: Jacky Snoep, Theresa Kouril
Submitter: Jacky Snoep
Purification of gluconeogenic enzymes from S. solfataricus in recombinant E.coli extracts
Creators: Jacky Snoep, Theresa Kouril
Submitter: Jacky Snoep
This assay uses a dual-wavelength spectrophotometer to quantify cytochromes present in the E. coli respiratory chain.
Creators: Alison Graham, Robert Poole, Jeff Green
Submitter: Alison Graham
This is a well-established, classical genetic method of constructing chromosomal monolysogenic fusions to a promoterless lacZ gene.
Creators: Alison Graham, Jeff Green, Robert Poole
Submitter: Alison Graham
The phosphorylation level of a particular protein can be determined using a procedure based upon western immunoblotting, with Phos-tag™ reagent present in the SDS-PAGE gel. The Phos-tag™ reagent, supplied in the form of Phos-tag™ acrylamide (Wako Pure Chemical Industries, AAL-107), causes proteins to be resolved both on the basis of size and phosphorylation state. This means that phosphorylated and de-phosphorylated forms of the same protein can be distinguished.
Creator: Matthew Rolfe
Submitter: Matthew Rolfe
The Gene-doctoring method of lambda-red deletion (Lee et al., 2009) was modified slightly to create chromosomal mutations of fnr.
Creator: Matthew Rolfe
Submitter: Matthew Rolfe
This method describes how one can quench metabolism of Escherichia coli and extract metabolites from many kinds of metabolite classes like: nucleotides, sugar-phosphates, organic acids ....
Creator: Stefan Stagge
Submitter: Stefan Stagge
Sample preparation procedure for metabolic analysis on T. b. brucei 427 using LC/MS
Creator: Dong-Hyun Kim
Submitter: Dong-Hyun Kim
Labelling and extraction procedure for uniformly 13C-labelled E. coli (MG1655) for absolute quantification using isotope dilution technique by LC-MS
Creator: Dong-Hyun Kim
Submitter: Dong-Hyun Kim
Sample preparation procedure for metabolic footprint analysis on T. b. brucei 427 using LC/MS
Creator: Dong-Hyun Kim
Submitter: Dong-Hyun Kim
Creator: Federico Rojas
Submitter: Federico Rojas
Assay methodologies for individual glycolytic isoenzymes from the Mendes Group, University of Manchester, UK
Creator: Hanan Messiha
Submitter: Walter Glaser
This is the general protocol for the glycolytic enzyme measurements. Detailed Information for each Enzyme can be found in the SOP: Assays for measuring the activities of the individual glycolytic isoenzymes of Saccharomyces cerevisiae
Creator: Hanan Messiha
Submitter: Walter Glaser
Creator: John Raedts
Submitter: John Raedts
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
Creator: Federico Rojas
Submitter: Federico Rojas
Protocol for transfer of plasmids into Clostridium acetobutylicum ATCC 824 by electroporation
Creators: None
Submitter: Ying Zhang
ClosTron mutants should always be subjected to Southern blot analysis to ensure that only one intron insertion has occurred.
Creators: Ying Zhang, Nigel Minton
Submitter: Ying Zhang
A protocol to improve conventional, recombination-based gene knock-out methodologies thtough the provision of negative selection markers, pyrE or codA.
Creators: Ying Zhang, Nigel Minton
Submitter: Ying Zhang
Protocol for transfer of plasmids into Clostridium spp. by conjugation
Creators: Ying Zhang, Nigel Minton
Submitter: Ying Zhang
A refined and streamlined procedure to generate mutant in a wide range of different clostridial species, using group II intron retargeting methodologies.
Creators: Ying Zhang, Nigel Minton
Submitter: Ying Zhang
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.
Creator: Matthew Rolfe
Submitter: Matthew Rolfe
This SOP describes the preparation of E. coli RNA
Creator: Matthew Rolfe
Submitter: Matthew Rolfe
Creator: Michael Kohlstedt
Submitter: Michael Kohlstedt
SOP for growing yeast in anaerobic conditions
Creator: Maksim Zakhartsev
Submitter: Maksim Zakhartsev