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Alkaline Phosphatase Treatment of Bacteriophage lambda Vector DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3976

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Removal of the 5'-phosphate groups from the internal termini of bacteriophage {lambda} arms is used to prevent self-ligation and suppress the background of nonrecombinant bacteriophages during cloning. - [Read Alkaline Phosphatase Treatment of Bacteriophage lambda Vector DNA Protocol]

Construction of Genomic DNA Libraries in Cosmid Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4000

Category: DNA Protocols: Cosmid Library Protocols

Protocol describes how to construct a library of 35-45-kb fragments of genomic DNA in the double cos site cosmid vector, SuperCos-1. The steps include: Linearization and dephosphorylation of SuperCos-1 DNA; Partial digestion of high-molecular-weight DNA with MboI; Dephosphorylation of high-molecular-weight genomic DNA; Ligation of cosmid arms to genomic DNA: Packaging and plating recombinants; Isolation and analysis of recombinant cosmids: Validation of the library. - [Read Construction of Genomic DNA Libraries in Cosmid Vectors Protocol]

Ligation of Bacteriophage lambda Arms to Fragments of Foreign Genomic DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3980

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Pilot ligations and packaging reactions are used to establish the amounts of fragmented genomic DNA and bacteriophage {lambda} arms that yield the maximum number of recombinants. Additional ligation and packaging reactions may then be set up to yield a comprehensive library of genomic DNA. - [Read Ligation of Bacteriophage lambda Arms to Fragments of Foreign Genomic DNA Protocol]

Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries - http://molecularstation.com/protocol-links/admin/dir_links_edit.php?action=N

Category: DNA Protocols: cDNA Library Protocols

The final stage of cDNA library construction involves optimizing ligation of the size-fractionated cDNA to bacteriophage {lambda} arms, followed by packaging and plating of the recombinant bacteriophages. - [Read Ligation of cDNA to Bacteriophage lambda Arms for the Construction of cDNA Libraries]

Preparation of Bacteriophage lambda DNA Cleaved with Two Restriction Enzymes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3987

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Many replacement vectors (e.g., the EMBL series, {lambda}2001, and {lambda}DASH) contain a series of restriction sites, arranged in opposite orientations, at each end of the central stuffer fragment. Digestion of these vectors with two different restriction enzymes yields left and right arms, a stuffer fragment, and short segments of the polycloning sites. These can easily be removed from the arms by differential precipitation with isopropanol or spun-column chromatography. - [Read Preparation of Bacteriophage lambda DNA Cleaved with Two Restriction Enzymes Protocol]

Purification of Bacteriophage lambda Arms: Centrifugation through Sucrose Density Gradients Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3990

Category: Viral Protocols

Protocol is used to prepare the arms of any bacteriophage {lambda} vector. - [Read Purification of Bacteriophage lambda Arms: Centrifugation through Sucrose Density Gradients Protocol]

Articles (1-1 of 1)

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.