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Simple, Reliable Steps for DNA Fragment Isolation and Purification Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4389

Category: Mouse Protocols: Mouse Genetics Protocols

Protocol describes a simple procedure for DNA fragment isolation and purification. The resulting DNA fragment can then. be used for microinjection to produce transgenic mice - [Read Simple, Reliable Steps for DNA Fragment Isolation and Purification Protocol]

Site-Specific Protein-DNA Photo-Cross-Linking: Analysis of Structural Organization of Protein-DNA - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4588

Category: DNA Protein Interactions Protocols

The multiprotein-DNA complex of interest is formed using the site-specifically derivatized DNA fragment. The complex is then UV-irradiated, initiating covalent cross-linking with proteins in direct physical proximity to the cross-linking agent. Extensive nuclease digestion is performed to eliminate uncross-linked DNA and convert cross-linked DNA to a cross-linked, radiolabeled nucleotide "tag." - [Read Site-Specific Protein-DNA Photo-Cross-Linking: Analysis of Structural Organization of Protein-DNA]

Subcloning of Yeast Artificial Chromosomes into Phage Lambda Protocol - http://humgen.wustl.edu/hdk_lab_manual/yeast/yeast9.html

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol for the subcloning of Yeast artificial chromosome into phage lambda. To subclone the large insert fragment of human DNA contained within a YAC into a bacteriophage lambda vector. The subclones are 15 to 23 kb in size, and can be used to identify new polymorphic markers from a known region of the genome, to map a specific locus, and/or to screen other libraries. - [Read Subcloning of Yeast Artificial Chromosomes into Phage Lambda Protocol]

Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4061

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

A synthetic oligonucleotide annealed to single-stranded DNA derived from a recombinant bacteriophage M13 or phagemid template is used to prime the synthesis of complementary radiolabeled DNA. Synthesis is catalyzed by the Klenow fragment of E. coli DNA polymerase I, which extends the annealed primer for various distances along the single-stranded template DNA. - [Read Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates Protocol]

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]

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Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Purifying DNA Fragments from Agarose Gels Alternate Protocol

An alternate protocol for purifying DNA fragments from agarose gels.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

Quick DNA Agarose Gel Electrophoresis Protocol.

A quick and simple protocol for the electrophoresis of DNA fragments using agarose gels.

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.

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.

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