Npromega t4 dna ligase pdf

Instructions for use of products m1801, m1804, m1794. For cohesive sticky ends, use 1 l of t4 dna ligase in a. Rapid ligation is based on the combination of t4 dna ligase with a unique 2x rapid ligation buffer. Therefore, invitrogen recommends the enzyme be kept at 20 c until within 510 minutes of use and returned immediately to 20 c after use. T4 dna ligase, 1 weiss ul 2x5 00 weiss u 10x t4 dna ligase buffer 1. T4 dna ligase, bluewhite cloning qualified protocolpdf 112 kb english. T4 dna ligase structure reveals a prototypical atpdependent. T4 dna ligase is provided with 10x reaction buffer. 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 cohesive or bluntended strands of dna between the 5. An overview of dna ligation, followed by a demonstration of the ligation of 2 dna fragments with t4 dna ligase for cloning purposes. T4 dna ligase can be used to join dna fragments with staggered or blunt ends. Ligation buffer for t4 dna ligase, 10 x conc 660 mm trishcl, 50 mm mgcl2, 50 mm dtt, 10 mm.

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 ligase dna ligation promega corporation. T4 dna ligase from multiple suppliers was tested in reac tions containing a fluorescent labeled single stranded, double stranded blunt, 3. The enzyme will not join singlestranded nucleic acids. T4 dna ligase, bluewhite cloning qualified protocol pdf 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 catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex dna or rna. T4 dna ligase is used in molecular cloning experiments to ligate join the. Bacteriophage t4 dna ligase is the first atpdependent ligase. Toll free in the usa 8003569526 telephone 6082744330 internet. Promega corporation 2800 woods hollow roadmadison, wi 537115399 u. 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. T4 dna ligase bluewhite cloning qualified protocol promega. For convenience, ligations may be done at room temperature 2025c.

T4 dna ligase catalyzes the joining of two strands of dna between the 5. Our crystal structure of t4 dna ligase bound to dna shows a. Promega enzyme resource guide 8 two ligases introduction dna ligases are primarily responsible for joining the gaps that form in dna during replication i. Description t4 dna ligase catalyzes the joining of two strands of dna between the 5.

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