Any other blunt or stickyend dna fragment can be cloned. The strataclone pcr cloning kit allows highefficiency, 5minute cloning of pcr products at room temperature, using the efficient dna rejoining activity of dna topoisomerase i and the dna. Thermo scientific clonejet pcr cloning kit for 20 reactions. Enhanced pcr cloning system pcrscript cloning kits. Strataclone vector mix, strataclone cloning buffer, strataclone control insert 5 ngl, solopack pcr cloning competent cells, puc18 control plasmid 0. Description high efficiency pcr cloning, features dna topoisomerase i and cre recombinase technologies be the first to write a. In the cloning process, the dna is removed from cells, manipulations.
The strataclone pcr cloning kit allows highefficiency, 5minute cloning of pcr products, using the efficient dna rejoining activity of dna topoisomerase i and the dna recombination activity of cre recombinase. No ligase, post pcr procedures, or pcr primers containing specific sequences are required. Sequence and ligase independent cloning slic as can be deduced from the name, ligase is not needed in the slic method. Lastly, if your pcr product is longer than usual i. Tim sanchez julie robinson mary buchanan stratagene. Map for the strataclone blunt pcr cloning vector pscbampkan 6. Note do not use taq dna polymerase, or enzyme blends containing predominantly taq dna polymerase, to amplify fragments for cloning using the strataclone blunt pcr cloning kit. The thermo scientific instaclone pcr cloning kit is a ta system for direct onestep cloning of pcr products with 3da overhangs 1. The high quality ta cloning vector ptz57rt is ready to use for efficient ligation with pcr products providing high cloning yields and low background. Qiagen pcr cloningplus kit transformation protocol.
Oct 24, 20 hello everyone, my first post on protocolonline. Box ii of the qiagen pcr cloningplus kit containing qiagen ez competent cells should be stored at 70c or. Our transformation protocol is often faster than the transformation of commercially available competent cells. My problem i cant seem to get a 2kb pcr amplicon cloned into the kits entry. Msds documents are not included with product shipments.
For descriptions of symbols on product labels or product documents, go to. The kit must produce 107 colonies from 250 ng of the exactstart rna control template when using epicentres transformax ec100 electrocompetent e. Nzya pcr cloning kit was designed to allow the direct cloning of pcr products with 5. Instaclone pcr cloning kit is a ta system for direct onestep cloning of pcr products with 3da overhangs generated by taq dna polymerase and other thermostable dna polymerases, which lack proofreading activity. Strataclone pcr cloning kit 5 solopack pcr cloning competent cells the provided solopack pcr cloning competent cells express cre recombinase, in order to circularize the linear dna molecules produced by topoisomerase imediated ligation.
The cells are provided in a convenient singletube transformation format. Strataclone pcr cloning kit 5 map for the strataclone pcr cloning vector pscaampkan feature nucleotide position. The clonejet pcr cloning kit is an advanced positive selection system for the highest efficiency cloning of pcr products generated with. Design of primers for pcr reaction for quadplex grna cloning the scalability of the multiplex grna cloning kit allows for. Almost everyone has heard of dolly, the cloned sheep born in 1996 but what about the. The high quality readytouse ta cloning vector ptz57rt was prepared by linearizing the ptz57r plasmid with eco32i and tailing with. The pcr script cloning kits are convenient for cloning pcr fragments generated with any pcr enzyme. The clonejet pcr cloning kit is an advanced positive selection system. The strataclone pcr cloning kit was used to clone pcr fragments from a common fluorescent protein 664 bp.
The qiagen pcr cloning kit and box i of the qiagen pcr cloningplus kit containing ligation master mix and pdrive cloning vector should be stored at 20c in a constanttemperature freezer or at 70c immediately upon receipt. The high quality ta cloning vector ptz57rt is ready to use for. The mighty tacloning kit allows for fast, easy cloning of pcr products with this ta cloning method. Protocols for cloning without restriction enzymes or. Map for the strataclone pcr cloning vector pscaampkan. The kit also allows direct cloning from amplification reactions without purification, and works well whether or not the primers used in the pcr possess 5. Thermo scientific clonejet pcr cloning kit, also available with dh10b cells k123120, is an advanced positive selection system for highefficiency cloning of. An introduction to pcr cloning pcr products generated using a nonproofreading dna polymerase which lacks 3 to 5 exonuclease activity, such as taq dna polymerase, are left with a single templateindependent nucleotide, deoxyadenosine a, at the 3. Limited label license for strataclone pcr cloning kits.
The kit must produce 107 colonies from 250 ng of the exactstart rna control template when using. These kits are available for both bluntend and ua cloning. Clonejet pcr cloning kit and are available as standalone strains or. Kit for sequencing fiveminute cloning of taq polymeraseamplified pcr products for sequencing catalog nos. The clonejet pcr cloning kit contains a novel, readytouse positive selection cloning vector pjet1. Ligationfree cloning highthroughput cloning seamless dna fusion multiple pcr fragment assembly uracil excisionbased cloning artificial microrna homologous recombination gateway. Protocol for exactstart fulllength cdna library cloning kit. Kit for sequencing version j 052 250276 topo ta cloning.
The exactstart fulllength cdna library cloning kit is functiontested in a control reaction. Fiveminute cloning of taq polymeraseamplified pcr products. The strataclone pcr cloning kit was used to clone pcr fragments from a common fluorescent protein 664 bp, gammaglobin 1. The cloning vector may be dna taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. The pdrive cloning vector contains the standard sequencing annealing sites for m forward and reverse, as well as t7 and sp6 primers. Transformation protocol for neb pcr cloning kit neb. Cloning of taq polymeraseamplified pcr products thermo.
Creator cloning kits the creator pdnr cloning kit cat. The dna insert can be readily excised from the versatile polylinker of ptz57rt, sequenced using standard mpuc primers or in vitro transcribed with t7 rna. Figure 1 in the qiagen pcr cloning kit handbook shows the pdrive. A overhangs, which result from amplifications using nonproofreading dna polymerases such as nzytaq dna polymerase. The neb pcr cloning kit contains a sufficient supply of materials to perform 20 x 10. Therefore, it takes only one hour from the completion of pcr to cell plating. A cloning vector is a small piece of dna that can be stably maintained in an organism, and into which a foreign dna fragment can be inserted for cloning purposes. Instaclone pcr cloning kit 30 rxns k1214 from thermo. Instaclone pcr cloning kit is a ta system for direct onestep cloning of pcr products with 3da overhangs generated by taq dna polymerase and other thermostable dna polymerases, which. An alternative to cloning, called the polymerase chain reaction pcr, can be used to directly amplify rare specific dna sequences in a complex mixture when the ends of the sequence are known.
Strataclone pcr cloning kit from agilent technologies. The cloning vector was prepared by cutting nzytech. The vector contains a lethal restriction enzyme gene that is disrupted by ligation of a dna insert into the cloning site. Cloning technology was invented during the twentieth century and now is poised to help define the twentyfirst. As a result, only bacterial cells with recombinant plasmids are able to form colonies. The donor vector in this kit, pdnr1r is designed to increase recovery of. Transformation protocol for neb pcr cloning kit the following protocol is designed for neb 10beta competent e. Taq dna polymerase catalyzes the nontemplate directed addition of an adenine residue to the 3. Primers are also provided, allowing screening for inserts by colony pcr andor sequencing. What sequencing primers can i use with the pdrive cloning. All components of the exactstart fulllength cdna library cloning kit are free of. Topo ta cloning provides a highly efficient, 5minute, onestep cloning strategy topo cloning for the direct insertion of taq polymeraseamplified pcr products into a plasmid vector. The pcrscript cloning kits are convenient for cloning pcr fragments generated with any pcr.
The strataclone pcr cloning kit allows highefficiency, 5minute cloning of pcr products at room temperature, using the efficient dna rejoining activity of dna topoisomerase i and the dna recombination activity of cre recombinase. K4575j10, k457501, k457540, k458001, k458040, k459501, k459540 a limited label license covers this product see purchaser notification. Choochoo cloning tm kits are highly efficient directional pcr cloning kits for rapid, ligase and restriction enzymeindependent cloning of pcr products. Ive got a problem with invitrogens pentr d topo cloning kit. In the cloning process, the dna is removed from cells, manipulations of the dna are carried out in a testtube, and the dna is subsequently put back into cells. The addition of a a by taq dna polymerase is not required and has no effect on cloning efficiency. Nov 08, 2018 lastly, if your pcr product is longer than usual i. Thermo scientific clonejet pcr cloning kit, also available with dh10b cells, is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna.
Cloning using the invitrogen ta cloning kit whats needed. Dna cloning cloning is the process of moving a gene from the chromosome it occurs in naturally to an autonomously replicating vector. The pcr fragments can be generated by taq dna polymerase or other high fidelity dna polymerases. Clonexpress ii one step cloning kit,rapid cloning,vazyme. Assemble ligation reactions using the chart below as a guide. Description the genscript cloneez pcr cloning kit is designed for the quick pcr cloning, it is especially powerful in highthroughput cloning of pcr products into any destination vector effectively by sidestepping the tedious and limiting tasks such as selecting proper restriction enzymes, phosphatases, or ligases. Limited label license for strataclone ultra blunt pcr cloning kits. Instaclone pcr cloning kit 30 rxns k1214 from thermo fisher. Clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna polymerase.