T4 dna ligase reaction buffer t4 dna ligase is the industry standard for performance and quality. There is considerable latitude in the temperature and time needed for successful ligations. T4 dna ligase catalyzes the formation of a phosphodiester linkage. The ligation reaction mixture can be used directly for bacterial transformation using conventional transformation procedures or with the thermo scientific. Fast ligation efficiency is equal to that obtained with t4 dna ligase in a standard 1 hour ligation. Tested for the absence of endo, exodeoxyribonucleases, ribonucleases and functionally tested in dna ligation. T4 dna ligase catalyzes the formation of phosphodiester bonds in the presence of atp between doublestranded dnas with 3. T4 dna ligase catalyzes the joining of two strands of dna between the 5phosphate and the 3hydroxyl groups of adjacent. T4 dna ligase recombinant form of the enzyme from t4 phage.
T4 dna ligase buffer contains atp, so repeated freeze thaw cycles can degrade atp, thereby decreasing the efficiency of ligation. Thermo scientific t4 dna ligase product help molecular. Ligation buffer optimized for fast and efficient dna ligation. T4 dna ligase catalyzes the joining of two cohesive or bluntended strands of dna between the 5. Invitrogen expresslink t4 dna ligase fisher scientific.
Thermo scientific t4 dna ligase product help posted in molecular cloning. Thermo scientific t4 dna ligase product help molecular cloning. T4 dna ligase 5 ul thermo scientific t4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl termini in duplex dna or rna. T4 dna ligase catalyzes the formation of a phosphodiester linkage between dna chains by condensing the 5phosphoryl group of one dna chain with the adjacent 3hydroxyl group of the other dna chain. Product information thermo scientific phusion site.
The 10x buffer has been tested for absence of endo, exodeoxyribonucleases. The universal buffer allows rapid single, double or multiple dna digestion within 515 minutes eliminating any need for buffer change or subsequent dna cleanup steps. The enzyme will not join singlestranded nucleic acids. T4 dna ligase catalyzes the formation of a phosphodiester bond between the terminal 5. T4 dna ligase buffer from thermo fisher scientific. A 1x concentration assures optimal enzyme activity. Rely on thermo scientific nalgene rapid flow filtration units. Thermo scientific t4 dna ligase is a single polypeptide. Browse a full range of enzymes inhibitors products from leading suppliers. Thermo scientific rapid dna ligation kit enables fast stickyend or bluntend dna ligation in only 5 minutes at room temperature. The efficiency of the fast ligation is analogous to that obtained with t4 dna ligase in a standard 1 hour ligation.
Introduction sitedirected mutagenesis is widely used in the study of. The unique t4 dna ligase buffer optimizes ligation, which can be performed in 5 minutes 1. Activity of dna modifying enzymes in fastdigest and. Tested for the absence of endo, exodeoxyribonucleases, ribonucleases and functionally dna ligation. Thermo scientific t4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl termini in duplex dna or rna.
Mix thoroughly, spin briefly and incubate 10 min at. This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex dna, rna or dna rna hybrids. If you are viewing this page as a nonregistered user, the. Fastdigest restriction enzymes thermo scientific 100 % buffer compatibility with all downstream applications common dna modifying enzymes are 100% active in fastdigest and fastdigest green buffer.
Dna modifying enzymes, such as klenow fragment, t4 dna ligase, alkaline phosphatases and t4 dna polymerase all have 100% activity in fastdigest buffer. Sinapse biotecnologia t4 dna ligase thermo scientific. Fastdigest restriction enzymes thermo scientific 100 % buffer compatibility with all downstream. 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 nucleic.
Understand the activity of dna modifying enzymes in cutsmart buffer and the required. One weiss unit is approximately equivalent to 22 enzymatics cohesive end units. Rapid dna ligation kit enables fast stickyend or bluntend dna ligation in only 5 minutes at room temperature. T4 dna ligase from thermo fisher scientific selectscience. Shop now at fisher scientific for all of your scientific needs. Y90001 5x t4 dna ligase buffer buffer composition 5x concentration1. Singlestranded nucleic acids are not substrates for thi. The 10x buffer has been tested for absence of endo, exodeoxyribonucleases, ribonucleases and functionally. Learn more about how this product is being used in the product citation tool. Tested for absence of endo, exodeoxyribonucleases, ribonucleases and functionally tested in dna ligation. Mix thoroughly, spin briefly and incubate 10 min at 22c.
Thermo scientific fastdigest bamhi fisher scientific. Our teams are in regular contact with suppliers and. Greetings i have been struggling to ligate a 555 bp insert into a 6. Thermo scientific t4 dna ligase, lc 1 ul t4 dna ligase.
It also joins dna fragments with either cohesive or blunt term. The kit contains t4 dna ligase and a speciallyformulated 5x rapid ligation buffer optimized for fast and efficient dna ligation. T4 dna ligase catalyzes the formation of phosphodiester bonds between neighboring 3. T4 dna ligase is the industry standard for performance and quality. Dna ligase is a specific type of enzyme, a ligase, ec 6. The enzyme used to ligate dna fragments is t4 dna ligase, which originates from the t4 bacteriophage. T4 dna ligase is provided with 10x reaction buffer.
Singlestranded nicks in doublestranded dna are also closed by t4 dna ligase. This products resides on a fisher scientific gsa or va contract. L of purified pcr product from the pcr cleanup plate, 2. The kit contains t4 dna ligase and a speciallyformulated 5x rapid ligation buffer optimized for fast and efficient dna l.
Active thermophilic dna polymerase still present in pcr mixture may alter the ends of the cleaved dna and reduce the ligation efficiency. Sep 17, 2012 ligation using t4 dna ligase amrita university. Contents t4 dna ligase, supplied with 10x concentrated ligation buffer that includes atp. The enzyme repairs singlestrand nicks in duplex dna, rna, or dnarna hybrids. The ligation reaction mixture can be used directly for bacterial. The 10x buffer has been tested for absence of endo, exodeoxyribonucleases, ribonucleases and functionally tested in dna ligation. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. The vector is further dephosphorylated with thermo scientifics fast ap to prevent. T4 dna ligase 200 units from thermo fisher scientific description t4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl. T4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl termini in duplex dna or rna.
T4 dna ligase requires atp, and if the buffer is very old, have been thawed. If you are viewing this page as a nonregistered user, the prices displayed is list price. Activity of dna modifying enzymes in cutsmart buffer. Highlights active in themo scientific restriction enzyme, pcr, and rt buffers when supplemented with atp. It plays a role in repairing singlestrand breaks in duplex dna in living organisms, but some forms such as dna ligase iv may specifically repair doublestrand breaks i. T4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex dna or rna. Thermo scientific rapid dna ligation kit fisher scientific. The enzyme efficiently joins blunt and cohesive ends and repairs singlestranded nicks in duplex dna, rna or dnarna hybrids. At a 1x concentration this reaction buffer assures optimal activity of the enzyme. Activity of dna modifying enzymes in cutsmart buffer neb. Thermo scientific fastdigest enzymes product information.
Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. The unique t4 dna ligase buffer optimizes ligation, which can be performed in 5 minutes. It is better to vortex or spin the t4 dna ligase enzyme. T4 dna ligase 200 units from thermo fisher scientific description t4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl termini in duplex dna or rna. Ligation using t4 dna ligase amrita university youtube. Thermo scientific t4 dna ligase, lc 1 ul t4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl termini in duplex dna or rna. It is recommended that the reaction buffer be discarded after one year of storage at 20c and replaced with fresh buffer to ensure maximum. It is commonly used to join dna molecules to each other.
What is supplied with the t4 dna ligase reaction buffer pack. The unique t4 dna ligase buffer optimizes ligation, which can be performed in 5 moreminutes. Cant clone cdna insert 797bp into pet22b with t4 dna ligase. The mixture of 10um forward primer with 5 end, 10x t4 dna ligase buffer, t4 pnk 10 units and nucleasefree water 1 incubated at 37c for 1 hour, and 2 incubated at 65c for 20 minutes to. T4 dna ligase 200 units from thermo fisher scientific.
L of ligation master mix to each well of a new pcr plate. The thermo scientific 10x t4 dna ligase buffer is used with t4 dna ligase. The enzyme repairs singlestrand nicks in duplex dna, rna, or dna rna hybrids. Singlestranded nucleic acids are not substrates for this enzyme. T4 dna ligase catalyzes the formation of phosphodiester bonds between doublestranded dna strands with 3 hydroxyl and 5 phosphate termini in the presence of atp. Use up to 5 l of the mixture to transform 50 l of chemically competent cells and 12 l per 50 l of electrocompetent cells. The kit contains t4 dna ligase and a speciallyformulated 5x. 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. Expresslink t4 dna ligase formulation is optimized for faster reaction times and more convenient incubation temperature than our other t4 dna ligase formulations. Intact genomics t4 dna ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5phosphate and 3hydroxyl termini in duplex dna or rna. What is the composition of the t4 dna ligase reaction. T4 dna ligase catalyzes the formation of phosphodiester bonds in the presence of atp between doublestranded dnas with 3 hydroxyl and 5 phosphate termini.
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