T4 dna ligase bluewhite cloning qualified protocol promega. T4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex dna or rna. Promega corporation 2800 woods hollow roadmadison, wi 537115399 u. Our crystal structure of t4 dna ligase bound to dna shows a. Dna ligase, which is expressed in the early stage of lytic development, has been implicated in t4 dna synthesis and genetic recombination. T4 dna ligase catalyzes the joining of two strands of dna between the 5. T4 dna ligase catalyzes the ligation of two rna substrates that are precisely aligned in a fully basepaired rnadna heteroduplex, whereas t4 rna ligase is used to join two singlestranded rnas in. The enzyme will not join singlestranded nucleic acids. For cohesive sticky ends, use 1 l of t4 dna ligase in a. Ligation buffer for t4 dna ligase, 10 x conc 660 mm trishcl, 50 mm mgcl2, 50 mm dtt, 10 mm. Rapid ligation is based on the combination of t4 dna ligase with a unique 2x rapid ligation buffer. Toll free in the usa 8003569526 telephone 6082744330 internet. T4 dna ligase can be used to join dna fragments with staggered or blunt ends. T4 dna ligase, bluewhite cloning qualified protocol pdf 112 kb english.
An overview of dna ligation, followed by a demonstration of the ligation of 2 dna fragments with t4 dna ligase for cloning purposes. Promega enzyme resource guide 8 two ligases introduction dna ligases are primarily responsible for joining the gaps that form in dna during replication i. T4 dna ligase, bluewhite cloning qualified protocolpdf 112 kb english. T4 dna ligase catalyzes the joining of two strands of dna between the 5phosphate and the 3hydroxyl groups of adjacent nucleotides in either a cohesiveended or bluntended configuration. T4 dna ligase structure reveals a prototypical atpdependent. The ligafast rapid dna ligation system is designed for the efficient ligation of cohesiveended dna inserts into plasmid vectors in just 5 minutes bluntended inserts in as little as 15 minutes. T4 dna ligase catalyzes the joining of two cohesive or bluntended strands of dna between the 5. Bacteriophage t4 dna ligase is the first atpdependent ligase. For convenience, ligations may be done at room temperature 2025c. Instructions for use of products m1801, m1804, m1794. T4 dna ligase is provided with 10x reaction buffer.557 420 127 973 573 1100 1044 964 1495 1270 984 200 878 1114 146 426 261 874 3 734 309 51 653 1562 1325 1056 1379 1287 278 796 1285 1113 317 702 1074 1399 1075 1213 241