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Promega t4 dna ligase manually

T4 DNA Ligase catalyzes the joining of two cohesive or bluntended strands of DNA between the 5phosphate and the 3hydroxyl groups of adjacent nucleotides.

The Promega t4 dna ligase manually t4 rna Unit Definition: 0. 01 Weiss unit of T4 DNA Ligase is the amount of enzyme required to catalyze the ligation of greater than 95 of 1g of HindIII fragments at 16C in 20 minutes. T4 DNA Ligase Joins two dsDNA strands with cohesive or blunt ends by using the 5phosphate and the 3hydroxyl groups of adjacent nucleotides. M1801, M1804, M1794 vector DNA 100ng insert DNA 17ng Ligase 10X Buffer 1l T4 DNA Ligase (Weiss units) 0. 11u NucleaseFree Water to final volume of 10l 2.

Incubate the reaction at: room temperature for 3 hours, or 4C overnight, or 15C for 418 hours. Notes: 1. There is considerable latitude in the temperature and time needed for successful ligations. T4 DNA Ligase catalyzes the joining of two strands of DNA between the 5'phosphate and the 3'hydroxyl groups of adjacent nucleotides in either a cohesiveended or bluntended configuration.

T4 PNK catalyzes the transfer of the gammaphosphate from ATP to the 5terminus of polynucleotides or to mononucleotides bearing a 5hydroxyl group. This product is available through the Promega Helix onsite stocking program.

We offer numerous convenient solutions to meet your lab's needs. T4 DNA Ligase. Unit Definition: 0. 01 Weiss unit of T4 DNA Ligase is the amount of enzyme required to catalyze the ligation of greater than 95 of 1g of HindIII fragments at 16C in 20 minutes.

When the concentrations of Pol I DNA polymerases are higher, it has an adverse effect on E. coli DNA ligase. T4. The DNA ligase from bacteriophage T4(Enterobacteria phage T4 is a bacteriophage that infects Escherichia coli bacteria. The T4 phage is a member of the Teven phages, a group including enterobacteriophages T2 and T6. T4 Polynucleotide Kinase catalyzes the transfer of the phosphate from ATP to the 5terminus of polynucleotides or to mononucleotides bearing a 5hydroxyl group.

The enzyme may be used to phosphorylate RNA, DNA and synthetic oligonucleotides prior to subsequent manipulations such as ligation and cloning. To dilute T4 DNA Ligase that will subsequently be stored at 20C, 50 glycerol storage buffer (Diluent Buffer A, NEB# B8001S) should be used; to dilute for immediate use, 1X Description: 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).

Rapid ligation is based on the combination of T4