After running the gel, compare the unligated lanes with the ligated lanes to verify evidence of activity. Ligafasttm rapid dna ligation system certificate of. Ligafast rapid dna ligation system protocolpdf promega. Rapid dna lib prep kitdna library preparation abclonal. The kit can be conveniently used for construction of pcrfree libraries from 100 ng of highquality fragmented genomic dna, or 10 ng of circulating cellfree dna cfdna. Litmus 28i vector was cut with either ecorv blunt or hindiii cohesive, treated with calf intestinal phosphatase and gel purified. The simplebiotech rapid dna ligation kit allows ligation of blunt or sticky ends in just 15 minutes. The thermostability enables multiple rounds of denaturation and ligation. The ligafast rapid dna ligation system is designed for the efficient ligation of stickyended dna inserts into plasmid vectors in just 5 minutes bluntended inserts in as little as 15 minutes. The kit includes t4 dna ligase and a specially formulated 5x rapid ligation buffer optimized for fast and efficient dna ligation. Depending on the dna concentration in the reaction, the ligation products will be either circular if the dna concentration is low or. Rapid dna ligation kit enables fast stickyend or bluntend dna ligation in only 5 minutes at room temperature. Fast ligation efficiency is equal to that obtained with t4 dna ligase in a standard 1 hour ligation.
Depending on the dna concentration, either circular low dna concentration or concatemeric high dna concentration ligation. Depending on the dna concentration in the reaction, the ligation products will be either circular if the dna concentration is low or concatemeric if the dna concentration is high. A rapid dna system is the collection of components that together performs a rapid dna analysis consisting of a rapid dna instrument, the pcr str typing kit rapid dna cartridge, and an integrated. A laboratory manual, cold spring harbor laboratory, ny 1989, 2nd ed. T4 dna ligase and buffer combination for rapid 5minute ligations. Rapid ligation is based on the combination of t4 dna ligase with a unique 2x rapid ligation buffer. Thermo scientific rapid dna ligation kit for 150 reactions. Using an optimized buffer system and t4 dna ligase, these kits allow ligation. Assemble the following reaction in a sterile microcentrifuge tube. The dna ligation kit, mighty mix is a 2x ligation premix for highefficiency ligations, especially for bluntended ligations, while the dna ligation kit, version 2. Dna ligat ion is an important technique in molecular cloning and in the generation of recombinant dna. I have been using this kit for the past 2 years for cloning inserts of 24 kb into plasmid vectors.
The rapid dna ligation kit enables fast stickyend or bluntend dna ligation in only 5 min at room temperature. Dna ligation is the act of joining together dna strands with covalent bonds with the aim of making new viable dna. This kit contains the new recombinant rapid alkaline phosphatase, the tool of choice for dephosphorylation of 5. Ligase master mix for the ligation of difficulttoligate dsdna fragments. Ligafast rapid dna ligation system promega corporation. Find and compare commercial and governmental sources for immunological and biological products using the linscotts directory search engine. The contents of the kit are ligation buffer, dna dilution buffer and t4 dna ligase. We offer a variety of ligation kits for joining nucleic acid fragments. Takaras dna ligation kits are simple, twocomponent systems that allow rapid performance of all cloning procedures requiring the covalent joining of dna molecules in vitro. The quick ligation kit enables ligation of cohesive end or blunt end dna fragments in 5 minutes at room temperature 25 c. Typical ligation reactions use 100200ng of vector dna. For rapid ligation of blunt ends, use t4 dna ligase, cat no. Cloning of fragments into either plasmid or phage vectors linker ligation recircularization of linear vector dna generation of libraries.
The selection of a dna ligation kit requires consideration of several factors. During cloning projects it is helpful to assess whether the ligation involves cloning a long insert, whether rapid ligation would aid. The efficiency of the fast ligation is analogous to that obtained with t4 dna. The rapid dna ligation kit is functionally tested in cloning experiment. The rapid dna ligation kit enables stickyend or bluntend dna ligation in only 5 minutes at room temperature see fig. The design of a cycled ligation assembly figure 1 takes advantage of a thermostable ligase specific for nicks in doublestranded dna e.
The substrates tested were prepared from short, synthetic oligonucleotides purchased from idt, and annealed using standard. This higher concentration is required for rapid ligation of blunt ends. Thermo scientific rapid dna ligation kit enables fast stickyend or bluntend dna ligation in only 5 minutes at room temperature. Joining of difficult to ligate dsdna fragments with blunt. The kit contains t4 dna ligase and a specially formulated 5x rapid ligation buffer optimized for fast and efficient dna ligation. The convenient kit enables ligation of dna in 5 minutes and dephosphorylation in 10 minutes.
The efficiency of this fast ligation is analogous to that obtained with t4 dna. Rapid dna ligation kit simplebiotech enzymes and more. Cloning, a laboratory manual, cold spring harbor laboratory. Fast ligation efficiency is equal to that obtained with t4 dna. The kit contains t4 dna ligase and a speciallyformulated 5x rapid ligation buffer optimized for fast and efficient dna ligation. Pcr primers provided in the kit are optional for users who prefer selfselected highfidelity dna. The rapid dna ligation kit can rapidly ligate dna with either blunt or sticky ends. Ligation sequencing kit rapid sequencing kit with transposase the transposase simultaneously cleaves template molecules and attaches tags to the cleaved ends rapid sequencing adapters are added to the tagged ends fragment lengths are a result of the random cleavage dna. The rapid dna ligation kit can rapi dly ligate dna with either blunt or sticky ends. The following ligation reaction of a 3kb vector and a 0. The ligafast rapid dna ligation system is designed for the.
Rapid ligation is based on the combination of t4 dna ligase with a unique 2x rapid ligation. The rapid dna ligation kit contains all reagents necessary for ligation. Avoid phenolchloroform extractions to remove alkaline phosphatase. The quick ligation kit enables ligation of cohesive end or blunt end dna fragments in 5 minutes at room temperature. Takaras dna ligation kit, mighty mix, is a onesolution premix ligation reagent that offers high efficiency ligations, particularly for bluntended ligation. Dna ligation kit, fast dna ligation kit, jetseq er and ligation kit, sigma dna ligation kit, roche rapid dna ligation kit. The ideal ratio between insert and vector in ligation reactions often has to be empirically determined. Rapid dna ligation kit california institute of technology. The kit allows for a small final reaction volume through the use of a single. What this product does number of tests the kit is designed for 40 or 160 dephosphorylation 1 g dna each and ligation reactions depending on pack size standard assay. Efficient adaptor ligation for the preparation of dsdna. What this product does number of tests 40 ligation reactions standard assay. Dephosphorylation requires 10 min, while ligation of dna is performed in a 5 min reaction.
Rapid dna ligation kit from roche applied science,the rapid dna ligation kit enables ligation of stickyend or bluntend dna fragments in just 5 min at 1525 c. Ligafast rapid dna ligation system protocolpdf 111 kb english. A rapid and simple method for dna engineering using cycled. The unopened kit is stable at 15 to 25c through the expiration date printed on the label. Conveniently perform your entire cloning reaction in the same tube. Manual, second edition, cold spring harbor laboratory press, cold spring harbor. I have compared the ligation efficiency of the rapid dna ligation kit to t4 dna. Kapa hyperplus kits are ideally suited for low and highthroughput ngs library construction workflows that require dna fragmentation, end repair, atailing, adapter ligation and library amplification optional. The dna ligation kit is a simple system that enables very rapid dna ligation reactions. 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. Thermo scientific rapid dna ligation kit from thermo.
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