User Panel

My Panel

Bookmark Science Articles

plasmid Protocols

Search results for: plasmid

Protocols » Search Results

Search Results Protocol Links Sort by: Hits | Alphabetical

DNA Ligation - http://www.bio.com/protocolstools/protocol.jhtml?id=p2053

Category: Molecular Biology Protocols: DNA Ligation Protocols

Description and protocol of the steps required to join together plasmid and insert fragments to create a new plasmid. Paul Kaufman, Univ California. - [Read DNA Ligation]

DNA Ligation Protocol from Takara Shuzo Co. - http://www.hawaii.edu/microbiology/MO/ligation.htm

Category: Molecular Biology Protocols: DNA Ligation Protocols

Insertion of DNA into Plasmid Vectors, Insertion of DNA into l Phage Vectors, Self-Circularization of Linear DNA (Intramolecular Ligation), Linker (Adaptor) Ligation. Alam's Lab Univ Hawaii. - [Read DNA Ligation Protocol from Takara Shuzo Co.]

DNA Plasmid Prep. Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D1.html

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

Protocol for DNA plasmid prep. - [Read DNA Plasmid Prep. Protocol]

DNA Preparation and Needle Loading for Gene Delivery by Microinjection Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4652

Category: Microinjection Protocols: Microinjection of DNA Protocols

For microinjection into cells, high-quality plasmid DNA is purified using standard procedures. The resulting preparation is then delivered into the microinjection needle by either a back-filling or a forward-filling approach. - [Read DNA Preparation and Needle Loading for Gene Delivery by Microinjection Protocol]

DNA Preparation, Plasmid DNA Alkaline Method Protocol - http://rothlab.ucdavis.edu/protocols/D10.html

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

Protocol for DNA preparation, plasmid DNA using alkaline method. - [Read DNA Preparation, Plasmid DNA Alkaline Method Protocol]

DNA Transfection Mediated by Lipofection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3870

Category: Transfection Protocols

Protocol should be viewed as a starting point for systematic optimization of transfection mediated by lipofecting agents. Once a positive signal has been obtained from a transfected plasmid carrying a standard reporter gene, optimal conditions for transfection can be established by systematic variation of parameters such as the initial cell density, the amount and purity of DNA, the media and serum, and the time of exposure of the cells to the cationic-lipid-DNA complex. - [Read DNA Transfection Mediated by Lipofection Protocol]

DNA Transfection Using Polybrene Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3875

Category: Transfection Protocols

Polybrene and DMSO can be used to achieve stable transformation of several types of cells by plasmid DNA. The yield of transformants is up to 15-fold greater with Polybrene than with calcium phosphate-DNA coprecipitation. However, there is no difference between the two methods in the efficiency of transformation of cells by high-molecular-weight DNA. - [Read DNA Transfection Using Polybrene Protocol]

Exon Trapping and Amplification for Construction of the Library Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4075

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol describes how to construct and amplify a genomic DNA library in a plasmid vector (pSPL3) equipped to express cloned exons in transfected mammalian cells. - [Read Exon Trapping and Amplification for Construction of the Library Protocol]

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]

Fabrication Protocol for DNA Microarrays - http://research.nhgri.nih.gov/microarray/fabrication.shtml

Category: DNA Microarray Protocols

This protocol describes the steps required to produce a cDNA microarray. Gene-specific DNA is produced by PCR amplification of purified template plasmid DNAs from cloned ESTs. The PCR product is purified by ethanol precipitation, thoroughly resuspended in - [Read Fabrication Protocol for DNA Microarrays]

Fluorescence In Situ Hybridization (FISH) on Human Chromosomes and Interphase Nuclei Protocol - http://users.ugent.be/~fspelema/neubla/protocol/fish.htm

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for fluorescence in situ hybridization (FISH) on human chromosomes and interphase nuclei. Includes: Plasmid probes; COSMID-, YAC- and LIBRARY PROBES; Probe detection. - [Read Fluorescence In Situ Hybridization (FISH) on Human Chromosomes and Interphase Nuclei Protocol]

Generation of Sets of Nested Deletion Mutants with Exonuclease III Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4078

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

The double-stranded DNA of recombinant plasmid, phagemid, or bacteriophage M13 replicative form DNA is digested with two restriction enzymes whose sites of cleavage both lie between one end of the target DNA and the binding site for universal primer. The enzyme that cleaves nearer the target sequence must generate either a blunt end or a recessed 3' terminus; the other enzyme must generate a four-nucleotide protruding 3' terminus. - [Read Generation of Sets of Nested Deletion Mutants with Exonuclease III Protocol]

High Efficiency FCC Preparation and Tx Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F4.html

Category: Competent Cells Protocols

This is an easy and straightforward protocol that gives efficiencies of 106 to 107 cfu/mg of plasmid DNA. A growth curve is required for each strain that is prepared.
- [Read High Efficiency FCC Preparation and Tx Protocol]

Hydoxylamine Mutagenesis of Plamid DNA Protocol - http://rothlab.ucdavis.edu/protocols/HA-mutagenesis.html

Category: Bacteria Genetics Protocols

Protocol for hydoxylamine mutagenesis of plasmid DNA. - [Read Hydoxylamine Mutagenesis of Plamid DNA 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]

Hydroxylamine Mutagenesis of Plasmid DNA Protocol - http://www-rcf.usc.edu/~forsburg/hydroxyl.html

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for hydroxylamine mutagenesis of plasmid DNA is ideal for random mutagenesis of plasmid DNA which is then used in a plasmid shuffle or screen for ts mutants. - [Read Hydroxylamine Mutagenesis of Plasmid DNA Protocol]

In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3813

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for in vitro mutagenesis using double-stranded DNA templates. Two oligonucleotides are used to prime DNA synthesis catalyzed by a high-fidelity thermostable polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and occupy the same starting and ending positions on opposite strands of the plasmid DNA. - [Read In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI]

In vitro RNA synthesis from plasmid-borne sequences under the control of T phage promoters - http://www.nottingham.ac.uk/~mbzspd/methods/In_vitro_RNA_synthes.html

Category: RNA Protocols: In Vitro Transcription T7 SP6 T3 Protocols

RNA, in vitro synthesis using T3 T7 promoters. Dr. Dawson Univ. Nottingham. - [Read In vitro RNA synthesis from plasmid-borne sequences under the control of T phage promoters]

Isolation of Arabidopsis cDNAs by Complementation in Yeast - http://www.arabidopsis.org/cshl-course/12-yeast.html

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Includes: Isolation of Arabidopsis LEU2 cDNAs by complementation of the yeast leu2 mutation; Recovery of plasmid DNA from yeast cells; Analysis of Arabidopsis cDNAs that complement a yeast leu2 mutation; Preparation of library DNA from the Lacroute cDNA library; Preparation of Yeast Media. - [Read Isolation of Arabidopsis cDNAs by Complementation in Yeast]

Isolation of BAC DNA from Small-scale Cultures Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4006

Category: BACs Bacterial Artificial Chromosome Protocols

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA. - [Read Isolation of BAC DNA from Small-scale Cultures 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]

Ligating Plasmid and Target DNAs in Low-melting-temperature Agarose Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3929

Category: Cloning Protocols

Protocol for ligating plasmid and target DNAs in low-melting-temperature agarose. Ligation in low-melting-temperature agarose is much less efficient than ligation with purified DNA in free solution and requires a large amount of DNA ligase. The method is used chiefly for rapid subcloning of segments of DNA in dephosphorylated vectors and assembling recombinant constructs. - [Read Ligating Plasmid and Target DNAs in Low-melting-temperature Agarose Protocol]

Long Term Storage of Bacterial Strains Protocol - http://humgen.wustl.edu/hdk_lab_manual/plasmid/plsmid02.html

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40% glycerol. - [Read Long Term Storage of Bacterial Strains Protocol]

Long-PCR Reagents and Guidelines - http://wheat.pw.usda.gov/~lazo/methods/lazo/longpcr.html

Category: PCR Protocols: Quantitative PCR Protocols

General guidelines for long-PCR conditions and enzyme mixtures. Efficient long-PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3' to 5' exo) is present at a lower concentration. Includes: For PCR with low-complexity templates (e.g., plasmid and cosmid inserts); For PCR with moderate-complexity templates (e.g., bacterial genomic DNA); For PCR with high-complexity templates (e.g., human genomic DNA). - [Read Long-PCR Reagents and Guidelines]

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]

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 26-50 out of 116 displayed.

First Previous 1 [2] 3 4 5 Next Last

Total records: 116