A read is counted each time someone views a publication summary such as the title, abstract, and list of authors, clicks on a figure, or views or downloads the fulltext. Yields of final ligation product for all reaction conditions using high concentration t4 dna ligase, the quick ligation kit and bluntta master mix. The enzyme has also been shown to catalyze the joining of rna to either a dna or rna strand in a duplex molecule, but will not join singlestranded. Dna ligase is a specific type of enzyme, a ligase, ec 6. Please refer to the product label for the intended use statements for specific products. T4 dna ligase catalyzes the joining of two cohesive or bluntended strands of dna between the 5. Pdf cloning and expression of a new bacteriophage shph. Reaction conditions 2x rapid ligation buffer t4 dna ligase 10x buffer 1 hour at 24c 259 29 16 hours at 4c 580 164 cloning efficiency was calculated as the number of white coloniesng vector dna from a ligation containing 8ng control insert dna 542bp and 50ng pgemt easy vector. T4 dna ligase catalyzes the templatedependent ligation of dna. Rapid ligation is based on the combination of t4 dna ligase with a unique 2x rapid ligation buffer. Promega corporation 2800 woods hollow roadmadison, wi 537115399 u. The enzyme will not join singlestranded nucleic acids. T4 rna ligase catalyzes the atpdependent ligation of singlestranded rna or dna onto the 5.
T4 dna ligase ligase dna ligation promega corporation. T4 dna ligase is provided with 10x reaction buffer. T4 dna ligase is used in molecular cloning experiments to ligate join the ends of cloned dna fragments, ligase is commonly to insert dna into a cloning vector. Ligafast rapid dna ligation system promega corporation.
Promega manufactures products for a number of intended uses. English canada download sdspdf 67 kb english united states. T4 dna ligase bluewhite cloning qualified protocol promega. Some engineering has been done to improve the in vitro activity of t4 dna ligase. Cloning and expression of a new bacteriophage shph dna ligase isolated from sewage. Description t4 dna ligase catalyzes the joining of two strands of dna between the 5. T4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex dna or rna. T4 dna ligase catalyzes the joining of two strands of dna between the 5. Canonical nucleosides can be utilized by t4 dna ligase as.