Cloning Protocols

Protocols and information related to cloning and expression. Including: Baculovirus Protein Expression; Mutagenesis; Site-Directed Mutagenesis; Vectors.

Protocols » Cloning Protocols

Cloning Protocols Categories

Baculovirus Protein Expression Protocols (6)

Vector Protocols (10)

Cloning Protocols Protocol Links Sort by: Hits | Alphabetical

Fragmentation of DNA by Sonication Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4538

Protocol describes a method for DNA fragmentation by sonication. During sonication, DNA samples are subjected to hydrodynamic shearing by exposure to brief periods of sonication. DNA that has been sonicated for excessive periods of time is extremely difficult to clone. - [Read Fragmentation of DNA by Sonication Protocol]

Fragmentation of DNA by Nebulization Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4539

Protocol describes a method for DNA fragmentation by nebulization, in which the fine mist created by forcing a DNA solution through a small hole in the nebulizer unit is collected. The size of the fragments obtained by nebulization is determined chiefly by the speed at which the DNA solution passes through the hole, altering the pressure of the gas blowing through the nebulizer, the viscosity of the solution, and the temperature. - [Read Fragmentation of DNA by Nebulization Protocol]

Ligation Optimization Protocol - http://www.fhcrc.org/science/labs/fero/Protocols/Ligation.html

Protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independently characterize the ligation kinetics of the vector and insert DNA fragments and then to combine them in optimal ratios. - [Read Ligation Optimization Protocol]

Ligation of DNA fragments Protocol - http://ruummc.med.uu.nl/subsite/RepHTML/Protocols/Ligation.html

Protocol for ligation of DNA fragments. - [Read Ligation of DNA fragments Protocol]

Attaching Adaptors to Protruding Termini Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3920

Adaptors are short double-stranded synthetic oligonucleotides that carry an internal restriction endonuclease recognition site and single-stranded tails at one or both ends. Adaptors are used to exchange restriction sites at the termini of linear DNA molecules. They may be purchased in phosphorylated and unphosphorylated forms. - [Read Attaching Adaptors to Protruding Termini Protocol]

Purification of PCR Products in Preparation for Cloning Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3825

Protocol describes how to use proteinase K to destroy thermostable enzymes and to purify amplified DNA in preparation for cloning. - [Read Purification of PCR Products in Preparation for Cloning Protocol]

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

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]