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Expression of F-tryptophan labelled protein - http://www.biochem.ucl.ac.uk/bsm/nmr/protocols/protocols/fluoexp.html

Category: Protein Protocols: Protein Expression Protocols

Expression of F-tryptophan labelled protein. Protocol can be adapted for Fluoro-phenylalanine or Fluro-tyrosine labelling. Transformation of competent E. coli cells for expression with plasmid. Dr. Chen, Dept. of Biochem. & Mol. Biology, University College, London. - [Read Expression of F-tryptophan labelled protein]

Expression of protein in minimal media - http://www.biochem.ucl.ac.uk/bsm/nmr/protocols/protocols/protexp.html

Category: Protein Protocols: Protein Expression Protocols

Expression of protein in minimal media using E. coli cells for expression. Dr. Chen, Biochem. & Mol. Biology, University College. London - [Read Expression of protein in minimal media]

Frozen Competent E. coli Preparation and Transformation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F3.html

Category: E Coli Protocols: E coli Transformation Protocols

Protocol for frozen competent E. coli preparation and transformation. - [Read Frozen Competent E. coli Preparation and Transformation Protocol]

Frozen Competent E. coli Preparation and Transformation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F3.html

Category: Competent Cells Protocols

This is a very easy protocol which yields transformation efficiencies appropriate for routine cloning and transformation. - [Read Frozen Competent E. coli Preparation and Transformation Protocol]

Frozen Competent E.Coli Cells - http://www.ciwemb.edu/labs/koshland/Protocols/BACTERIA/ecolicells.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Frozen Competent E.Coli Cells. Methods and protocol for freezing E.Coli Bacteria Competent Cells in aliquots for later use re-use. (Inoue et al., 1990 Gene 96:23). Koshland. - [Read Frozen Competent E.Coli Cells]

Generation of Gene Deletions and Gene Replacements in Escherichia coli Protocol - http://www.biologicalprocedures.com/bpo/arts/1/123/m123.pdf

Category: E Coli Protocols: E coli Genetics Protocols

Protocol for the generation of gene deletions and gene replacements in Escherichia coli O157:H7 using a temperature sensitive allelic exchange system. Technology requires flanking DNA to be cloned into a temperature sensitive vector but the resulting clone allows great flexibility for further modification of the target sequence. It is therefore highly suited to the study of genes in which several rounds of changes are envisaged. - [Read Generation of Gene Deletions and Gene Replacements in Escherichia coli Protocol]

Hydroxylamine Mutagenesis of Plasmid DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4156

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Protocol produces mutagenized DNA that can be used for transformation of either Escherichia coli or yeast. - [Read Hydroxylamine Mutagenesis of Plasmid DNA Protocol]

Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes - http://omrf.ouhsc.edu/~frank/GT11.html

Category: RNA Protocols: cDNA Libraries Library

Adsroption of Anti-E. coli Antibodies, cDNA Library Screening, Frank Lab. Oklahoma Medical Research Foundation - [Read Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes]

Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4317

Category: C. elegans Protocols: C. elegans Genetics Protocols

Protocol describes an easily scalable way of introducing double-stranded RNA (dsRNA) in Caenorhabditis elegans: feeding the nematode with bacteria that express dsRNA. When using an RNase-III-negative Escherichia coli strain (HT115), the efficiency of this method is comparable to the alternative. - [Read Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol]

Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4317

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol describes an easily scalable way of introducing double-stranded RNA (dsRNA) in Caenorhabditis elegans: feeding the nematode with bacteria that express dsRNA. When using an Rnase-III-negative Escherichia coli strain (HT115), the efficiency of this method is comparable to the alternative. - [Read Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol]

Isolation of Plasmid DNA from Yeast Cells: A Ten-Minute Preparation - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4150

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast DNA Isolation Protocols

Protocol describes the purification release of plasmid DNA from yeast cells. The resulting plasmid DNA can be used to transform Escherichia coli or yeast. - [Read Isolation of Plasmid DNA from Yeast Cells: A Ten-Minute Preparation]

Isolation of Total RNA form E. coli for Microarrays Protocol - http://bugarrays.stanford.edu/protocols/rna/Total_RNA_from_Ecoli.pdf

Category: E Coli Protocols: E coli Nucleic Acid Protocols

Protocol for the isolation of total RNA from E. coli for Microarrays. Includes: Harvesting total RNA; DNase I treatment of RNA samples. - [Read Isolation of Total RNA form E. coli for Microarrays Protocol]

Isolation of Total RNA From E. Coli for Microarrays Protocol - http://bugarrays.stanford.edu/protocols/rna/Total_RNA_from_Ecoli.pdf

Category: Microarray Protocols: Microarray Analysis Protocols

Protocol for isolation of total RNA from E. coli for microarrays. Includes: Harvesting total RNA; Dnase I treatment of RNA samples. - [Read Isolation of Total RNA From E. Coli for Microarrays Protocol]

Ligation with GUS vector and Transformation - http://www.owlnet.rice.edu/~bios311/bios311/bios312/312day3.html

Category: Molecular Biology Protocols: DNA Ligation Protocols

Ligation with GUS vector and Transformation. Chemical Transformation of E. coli. Ligation Procedure for LigaFastâ„¢ Rapid DNA Ligation System (Promega). B. Beason, Rice University. - [Read Ligation with GUS vector and Transformation]

Maintenance of Probes in Bacteria Including Escherichia coli Protocol - http://wheat.pw.usda.gov/~lazo/methods/lazo/bactstor.html

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on Luria-Bertani (LB) agar on or in LB broth containing Ampicillin (30 µg/ml) or Carbenicillin (50 µg/ml broth, 100 µg/ml agar). E. coli strains are usually preserved in stab agar or glycerol for mid-term storage and lyophilized for long-term storage. - [Read Maintenance of Probes in Bacteria Including Escherichia coli Protocol]

Mini-Prep Protocol - http://ivaan.com/protocols/107.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Protocol for mini-prep. Includes: PREPARATION OF E. COLI; PRODUCTION OF CLEAR LYSATE; cDNA ISOLATION. - [Read Mini-Prep Protocol]

Molecular and Genetic Toxicology Studies on Microbial Mutagenesis - http://www.sri.com/biosciences/bdd/toxic/protocol.html#4

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies on microbial mutagenesis. Includes: G101 Salmonella/Microsome Plate Incorporation Assay (Ames Test), G101-R, G102 Salmonella/Microsome Preincubation Assay (Ames Test), G102-R, G103 Salmonella/E. Coli Reverse Mutation Assay (using four Salmonella and one E. coli strains), G104 Salmonella/E. Coli Reverse Mutation Assay (using four Salmonella and two E. coli strains), G105, G106. - [Read Molecular and Genetic Toxicology Studies on Microbial Mutagenesis]

Moving E. coli Genes into Salmonella Protocol - http://rothlab.ucdavis.edu/protocols/eco-genes-into-sty.html

Category: E Coli Protocols: E coli Genetics Protocols

Protocol for moving E. coli genes into Salmonella. - [Read Moving E. coli Genes into Salmonella Protocol]

Plating Bacteriophage M13 Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3993

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Bacteriophage M13 forms turbid plaques on lawns of male strains of E. coli. - [Read Plating Bacteriophage M13 Protocol]

Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in E coli Protocol - http://www.plantmethods.com/content/1/1/6

Category: E Coli Protocols: E coli Genetics Protocols

Protocol for precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli. Describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in E coli Protocol]

Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli - http://www.plantmethods.com/content/1/1/6

Category: Plant Biology Protocols: Plant Genetics Protocols

Describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli]

Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli - http://www.plantmethods.com/content/1/1/6

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli. Includes: Key steps in the EL250 RED-HR locus rescue and engineering procedure; Primer design and plasmid constructs; AtSTM gap-repair construct; Targeting construct backbone; Preparation of electrocompetent EL250 cells; Transformation of BAC F24o1 and induction of recombinogenic function in EL250; AtSTM locus rescue from BAC F24o1 by gap-repair HR. - [Read Precision Engineering of Plant Gene Loci by Homologous Recombination Cloning in Escherichia Coli]

Preparation and High-efficiency Transformation of Competent E. coli - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3942

Category: Bacteria Transformation Protocols

Protocol generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). - [Read Preparation and High-efficiency Transformation of Competent E. coli]

Preparation and Transformation of Competent E. coli Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3942

Category: E Coli Protocols: E coli Transformation Protocols

Procedure generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). IMPORTANT All steps in this protocol should be carried out aseptically. - [Read Preparation and Transformation of Competent E. coli Protocol]

Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3932

Category: Bacteria Transformation Protocols

Protocol used to prepare batches of competent bacteria that yield 5 x 106 to 2 x 107 transformed colonies/µg of supercoiled plasmid DNA. - [Read Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol]

Articles (1-2 of 2)

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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