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Bidirectional Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4139

Category: PCR Protocols: PCR Cloning Protocols

Protocol is for bidirectional, blunt-end cloning of DNA fragments. The target DNA is PCR amplified and 3'-extensions are polished with Pfu DNA polymerase. The amplicon is ligated to a blunt-ended plasmid DNA, and the products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction to relinearize any self-religating vector DNA. - [Read Bidirectional Cloning of PCR Products Protocol]

C. elegans RNAi Protocol - http://www.zoology.ubc.ca/~alorch/rnai-protocol.htm

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

Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding. - [Read C. elegans RNAi Protocol]

CaCl2 Competent Cell Stock Protocol - http://www.bio.indiana.edu/~chenlab/potocols/CompetentCellStock.htm

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Simple Protocol for CaCl2 Competent Cells. Chen Lab. Indiana University Bloomington - [Read CaCl2 Competent Cell Stock Protocol]

Competent cell preparation - http://www.chem.uga.edu/scottgrp/GrpProtocols/Competent_cell_preparation.htm

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Bacterial E.Coli competent cell preparation. Calcium Chloride Protocol, Preparation of calcium chloride competent cells for frozen storage, Electroporation Protocol, Electroporation Protocol for transformations using double-stranded plasmids, Reference: - [Read Competent cell preparation]

Competent Cell Preparation [CaCl2] - http://www.cf.ac.uk/biosi/staff/kille/Methods/Gene/Comp_cell_CaCl.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Nice protocol for the preparation of Bacterial cells to allow uptake of plasmid vectors. Dr Peter Kille. Hot Metals Cardiff. - [Read Competent Cell Preparation [CaCl2]]

Competent cells, calcium chloride method, E. coli, description - http://web.uct.ac.za/depts/mmi/bbhelp/bac1.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Competent cells, calcium chloride method, E. coli, description. CaCl2 and DMSO methods for preparing bacterial and E.Coli competent cells. Zappe, H. BBHelp. - [Read Competent cells, calcium chloride method, E. coli, description]

Directional Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4140

Category: PCR Protocols: PCR Cloning Protocols

Protocol is for directional blunt-end cloning of DNA fragments. The target DNA is PCR-amplified, 3'-extensions are polished with Pfu DNA polymerase, and the amplicon is ligated to a blunt-ended plasmid DNA. The products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction to relinearize any self-religating vector DNA. - [Read Directional Cloning of PCR Products Protocol]

Electro Competent Cells Protocol - http://hulab.tamu.edu/wiki/index.php/Electro_competent_cells

Category: E Coli Protocols: E coli Transformation Protocols

Protocol for electro competent cells. - [Read Electro Competent Cells Protocol]

Electro Competent Cells Protocol - http://hulab.tamu.edu/wiki/index.php/Electro_competent_cells

Category: Competent Cells Protocols

Protocol for electro competent cells. Preparation of Electro competent E. coli cells (per the BioRad directions). - [Read Electro Competent Cells Protocol]

EMBL Cloning protocols - http://www.embl-hamburg.de/display?file=~geerlof/webPP/protocoldb/Cloning/pdb_cloning.html

Category: DNA Protocols: DNA Cloning

Preparation of oligo solutions PCR experiments, Digestion of insert DNA, Digestion and dephosphorylation of vector DNA, Ligation of DNA fragments with sticky ends, Ligation of DNA fragments with blunt ends, Preparation of chemically competent E. coli - [Read EMBL Cloning protocols]

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]

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]

Frozen Yeast Transformation Protocol - http://www.umanitoba.ca/faculties/medicine/biochem/gietz/Frozen.html

Category: Yeast Protocols

This Protocol allows You to prepare Frozen Yeast cells that are competent for transformation after thawing. - [Read Frozen Yeast Transformation Protocol]

Hanahan Competent Cell Protocol. - http://www.rockefeller.edu/labheads/vosshall/protocols/Hanahan.pdf

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Hanahan Competent Cell Protocol. PROTOCOL PRODUCES ABOUT 160 X 250ul ALIQUOTS OF COMPETENT CELLS. Vosshall. Rockefeller. - [Read Hanahan Competent Cell Protocol.]

How to Make Competent Cells - Calcium Chloride - http://sosnick.uchicago.edu/competent_cells.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

How to Make Competent Cells - Calcium Chloride. Sosnick Lab, Univ. Chicago - [Read How to Make Competent Cells - Calcium Chloride]

Method: Preparation of Competent Cells - http://humgen.wustl.edu/hdk_lab_manual/plasmid/plsmid13.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Detailed protocol for creating bacterial competent cells preparation. Matthew S. Holt - [Read Method: Preparation of Competent Cells]

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]

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

Category: E Coli Protocols: E coli Transformation Protocols

Protocol is is 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]

Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3944

Category: Bacteria Transformation Protocols

Protocol reproducibly generates competent cultures of E. coli that yield 1 x 108 to 3 x 108 transformed colonies/µg of plasmid DNA. The protocol works optimally when the bacterial culture is grown at 18°C. If a suitable incubator is not available, a standard bacterial shaker can be set up in a 4°C cold room and regulated to 18°C. - [Read Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol]

Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3944

Category: E Coli Protocols: E coli Transformation Protocols

Preparation of Competent Bacterial Cells - http://www.cbs.umn.edu/~amundsen/chlamy/methods/competent.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Simple E.coli competent cell Protocol. Based on a protocol from Neil Olszewski and Scott Medberry.. Craig D. Amundsen - [Read Preparation of Competent Bacterial Cells]

Articles (1-3 of 3)

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

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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