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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]

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]

How to Transform Neurospora crassa by Electroporation - http://www.fgsc.net/neurosporaprotocols/How%20to%20transform%20Nc%20by%20electroporation.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to transform Neurospora crassa by electroporation. - [Read How to Transform Neurospora crassa by Electroporation]

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]

LiOAc Yeast Transformation Protocol - http://proteome.wayne.edu/transform.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

LiOAc/TE, 40% PEG in LiOAc/TE. Transformation Method and Protocol for Yeast. Finley Lab, Wayne State. - [Read LiOAc Yeast Transformation Protocol]

LiOAc Yeast Transformation Protocol - http://proteome.wayne.edu/transform.html

Category: Yeast Protocols: Yeast Transformation Protocols

Protocol for LiOAc Yeast transformation. - [Read LiOAc Yeast Transformation Protocol]

PCR-Mediated Gene Disruption: One-Step Method Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4169

Category: PCR Protocols

DNA prepared by PCR-mediated gene disruption can be used to transform yeast in gene replacement experiments. This protocol uses two primers, tailed with approximately 50 nucleotides homologous to the gene of interest, that target insertion of the PCR product to that locus. Each primer ends with a universal sequence that is designed to amplify various selectable markers from plasmid templates. - [Read PCR-Mediated Gene Disruption: One-Step Method Protocol]

Protein Expression and Purification Protocol - http://sosnick.uchicago.edu/Protein_Express&Purify.html

Category: Recombinant Protein Protocols

From plasmid to protein using bacterial expression. Transform appropriate DNA plasmid, Make a starter culture for protein expression, bacteria culture for protein expression. Sosnick Group Chicago. - [Read Protein Expression and Purification Protocol]

Working with Bacterial Artificial Chromosomes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4010

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol describes how to transform E. coli with BAC DNA by electroporation. - [Read Working with Bacterial Artificial Chromosomes Protocol]

Articles (1-4 of 4)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

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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