User Panel

My Panel

Bookmark Science Articles

plasmid Protocols

Search results for: plasmid

Protocols » Search Results

Search Results Protocol Links Sort by: Hits | Alphabetical

Synthesis of dsRNA for RNAi in Drosophila: Plasmid Template Method Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4512

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

Protocol describes how to prepare double-stranded RNA (dsRNA) for RNA interference in Drosophila by synthesis of individual RNA strands from linearized plasmid templates, followed by annealing of the strands. DsRNA molecules with a length of 500-800 bp seem to be most active. The dsRNA can be made from cDNA or genomic DNA templates, as long as most of the dsRNA corresponds to presumptive exon sequence. - [Read Synthesis of dsRNA for RNAi in Drosophila: Plasmid Template Method Protocol]

Tetracycline as Regulator of Inducible Gene Expression Protocol II - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3957

Category: Transfection Protocols: Stable Transfection Protocols

This stage of the procedure describes the transfection with target genes of cell lines already expressing inducible tTA. In this example, the target genes are transfected on a plasmid that carries puromycin resistance as a selectable marker. - [Read Tetracycline as Regulator of Inducible Gene Expression Protocol II]

Topo Cloning Procedure - http://preuss.bsd.uchicago.edu/protocols/topo.html

Category: DNA Protocols: TOPO Cloning

Topo Cloning procedure for cloning of small amounts DNA fragments by PCR. Cloning the PCR fragment into the TOPO vector, transforming E. Coli cells, and using blue/white selection to determine transformants. Culture inoculation, Qiagen Plasmid Prep protocol for plasmid isolation of the plasmid containing the cloned copies. Preuss Lab, Univ. Chicago. - [Read Topo Cloning Procedure]

Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4664

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. The calcium phosphate method can be used to transfect neurons that have already been growing on coverslips in vitro. Transfected cells suitable for imaging can be obtained in cultures up to 15 days in vitro. - [Read Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid]

Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4664

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols

Protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. - [Read Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid Protocol]

Transfection of Hippocampal Neurons with Plasmid DNA Using Calcium Phosphate Coprecipitation - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4445

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This protocol describes transfection of plasmid DNA into primary hippocampal neurons using DNA/calcium-phosphate (CaPO4) coprecipitation. The precise pH of the transfection medium and the incubation time of cells with the coprecipitate are critical for reproducible and efficient transfection. Once these parameters are optimized for a given plasmid, the method is easily adapted for transfection of other established cell lines. - [Read Transfection of Hippocampal Neurons with Plasmid DNA Using Calcium Phosphate Coprecipitation]

Transformation of Agrobacterium Using Electroporation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4665

Category: Bacteria Transformation Protocols

Protocol describes a method for transforming Agrobacterium with plasmid DNA using electroporation in a manner similar to that commonly used for Escherichia coli. Although the transformation efficiency for Agrobacterium is lower than that for E. coli, it is possible to obtain adequate numbers of Agrobacterium transformants with this technique. - [Read Transformation of Agrobacterium Using Electroporation Protocol]

Transformation of Agrobacterium Using the Freeze-Thaw Method Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4666

Category: Bacteria Transformation Protocols

Protocol describes a method for transforming Agrobacterium with plasmid DNA using a freeze-thaw technique. Although the transformation efficiency for Agrobacterium is lower than that for Escherichia coli, it is possible to obtain adequate numbers of transformants with this technique. - [Read Transformation of Agrobacterium Using the Freeze-Thaw Method Protocol]

Transposon Mediated Mutagenesis Protocol - http://www.genome.wisc.edu/resources/protocols/tnmutationprotocol.htm

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for transposon mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid. - [Read Transposon Mediated Mutagenesis Protocol]

Transposon-Mediated Mutagenesis Protocol - http://www.genome.wisc.edu/resources/protocols/tnmutationprotocol.htm

Category: Bacteria Genetics Protocols

Protocol for transposon-mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid. - [Read Transposon-Mediated Mutagenesis Protocol]

Two Hybrid System Transformation Protocol - http://www.umanitoba.ca/faculties/medicine/biochem/gietz/2HS.html

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

High Efficiency Transformation of a yeast strain requiring maintenance of a plasmid. - [Read Two Hybrid System Transformation Protocol]

Two-hybrid Systems Stage 1: Characterization of a Bait-LexA Fusion Protein Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3895

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol describes how to generate a plasmid construct (pBAIT) that expresses a target protein fused to the bacterial LexA protein. PBAIT is cotransformed into yeast with a lexAop-lacZ reporter plasmid carrying the bacterial lacZ gene under the control of the lexA operator. The recipient yeast strain contains a chromosomally integrated leu2 reporter gene, also under the control of the lexA operator. - [Read Two-hybrid Systems Stage 1: Characterization of a Bait-LexA Fusion Protein Protocol]

Two-hybrid Systems Stage 2: Selecting an Interactor Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3888

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

In this stage of the protocol, a mammalian cDNA library constructed in a plasmid such as pJG4-5 is transformed into yeast strains containing pBAIT and the lexAop-lacZ reporter plasmid. PJG4-5 expresses the cloned cDNAs from a cassette containing a transcriptional activation domain and other moieties under the control of the yeast GAL1 promoter. - [Read Two-hybrid Systems Stage 2: Selecting an Interactor Protocol]

Use of PCR for Quality Control of a Peptide DNA Library Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4137

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Protocol describes the use of PCR to characterize a peptide library encoded in a plasmid vector. In this example, the library was obtained by transforming bacteria with the ligation reaction at the end of Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides. - [Read Use of PCR for Quality Control of a Peptide DNA Library Protocol]

Vectors and Agrobacterium Hosts for Arabidopsis Transformation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.ip29

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

A. tumefaciens is a soil-dwelling bacterium that transforms normal plant cells into tumor-forming cells by inserting a piece of bacterial DNA (the transfer, or "T," DNA) into the plant cell genome. The Ti plasmid also carries many of the transfer functions for mobilizing the T-DNA. This article provides a brief discussion of the principles of T-DNA transformation, including consideration of T-DNA vectors and their hosts. - [Read Vectors and Agrobacterium Hosts for Arabidopsis Transformation Protocol]

Yeast Miniprep (Plasmid Rescue) Protocol - http://www.biochem.emory.edu/labs/acorbe2/yeastrescue.html

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

Protocol for Yeast miniprep (plasmid rescue). - [Read Yeast Miniprep (Plasmid Rescue) Protocol]

Articles (1-6 of 6)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

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.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

[1-6]

Items 101-116 out of 116 displayed.

First Previous 1 2 3 4 [5] Last

Total records: 116