This kit utilizes the 3adenosine overhangs produced during amplification. Samples with more than one clade on rflp screening or with additional unidentified bands were cloned using the clonejet pcr cloning kit thermo scientific, waltham, ma, usa with competent e. Dec 08, 2014 the thermo scientific clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna polymerase. For qiagen pcr cloning kit qiagen pcr cloningplus kit pdf 771kb english format. The strataclone pcr cloning kit is designed for efficient cloning of pcr fragments. The thermo scientific instaclone pcr cloning kit is a ta system for direct onestep cloning of pcr products with 3da overhangs 1. The kit works well with both phosphorylated and nonphosphorylated dna fragments. The pcr fragment was cloned subsequently using the clonejet pcr cloning kit fermentas. Followed thermo scientific clonejet pcr bluntend cloning protocol.
Jan 19, 2015 for sequence validation, the pcr product was subcloned using clonejet pcr cloning kit thermo scientific. The high quality ta cloning vector ptz57rt is ready. 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. For qiagen pcr cloning kit qiagen pcr cloningplus kit pdf 771kb english format file size language download get adobe reader contact qiagen. Any other blunt or stickyend dna fragment can be cloned. Transformation analysis of recombinant clones the clonejet pcr c loning kit is compatible with all common li laboratory strains. Pdf reproducibility of vibrionaceae population structure in. Pcr and rtpcr products corresponding to mock or advtransduced myoblasts were purified by using the jetquick gel extraction spin kit genomed, according to the. Gelanalyze the pcr product for specificity and yield before cloning. For convenience in mapping and manipulation of the insert, the pjet1. Mar 30, 2010 pcr cloning procedure consisting of the four following steps. Ligation times longer than 30 min are not recommended and may decrease cloning efficiency.
Purified amplicons were then subcloned using the clonejet pcr cloning kit thermofisher scientific, and subsequently transformed into. The clonejet pcr cloning kit is an advanced positive selection system. Specific pcr products of may 01, 2019 the clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products. Strataclone pcr cloning kit from stratagene biocompare. The clonejet pcr cloning kit contains a novel, readytouse positive selection cloning vector pjet1. Hi, i wanted to ask, how can i release an inserted dna into pjet1. Clonejet pcr cloning kit k1231 from thermo fisher scientific. The clonejet pcr cloning kit is an advanced positive selection system for the highest. Cloning kits clonejet pcr cloning kit k1231 20 rxns k1232 40 rxns rapid dna ligation kit k1422 50 rxns k1423 150 rxns fast dna end repair kit k0771 50 rxns alicator lic cloning. The vector contains a lethal restriction enzyme gene that is disrupted by ligation of a dna insert into the cloning site. The agilent pcmvscript pcr cloning kit is a polymerase chain reaction pcr cloning method that can be performed in 1 hour without adding bases to the primers. Pcr cloning procedure consisting of the four following steps. Any other blunt or stickyend dna fragment can be cloned as well.
Clonejet pcr cloning kit with dh10b competent cells. Choochoo cloning tm kits are highly efficient directional pcr cloning kits for rapid, ligase and restriction enzymeindependent cloning of pcr products. It is ideal for phosphorylated or nonphosphorylated dna fragments. Combine the following reagents sequentially on ice. An 8 l aliquot of the recovered pcr sample was ligated into pjet1. Order toll free 800 4245444 technical services 800 8944. Thermo scientific clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any.
The vector contains a lethal restriction enzyme gene that is disrupted by ligation. The kit contains a positive selection vector ensuring that only bacterial cells containing recombinant plasmids are able to form colonies and eliminating the need for expensive bluewhite screening. The kit contains a positive selection vector ensuring that only bacterial cells. See attached the thermo scientific clonejet pcr cloning kit. When i cloned pcr products in pjet i plate something like 2550 l of the transformation and the plates are full with colonies. Clonejet an advanced positive selection system for high. Pcrscript amp cloning kit 3 introduction the stratagene pcrscript amp cloning kit is a polymerase chain reaction pcr cloning method that can be performed in 1 hour without. Specific pcr products of 3 kb, ligation can be prolonged to 30 min. Samples with more than one clade on rflp screening or with additional unidentified bands were cloned using the clonejet pcr cloning kit thermo scientific, waltham, ma, usa with. May 03, 2019 the pcr fragment was cloned subsequently using the clonejet pcr cloning kit fermentas. The thermo scientific clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna. The kit contains strataclone vector mix, strataclone cloning buffer, solopack pcr cloning competent cells, strataclone control insert and puc18 control plasmid. The cells are provided in a convenient singletube transformation format. The neb pcr cloning kit contains a sufficient supply of materials to perform 20 x 10.
Sixty independent clones were analyzed by colony pcr using the smartiiaprimer and the genespecific gldpa race6 oligonucleotide 5. As a result, only bacterial cells with recombinant plasmids are able to form colonies. The traditional path for orf cloning has usually started with experimental observations driving the identification of one or several genes of interest to a. Primers are also provided, allowing screening for inserts by colony pcr andor. 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 polymerase. Primers are also provided, allowing screening for inserts by colony pcr andor sequencing. According to kit self ligated vector should not grow. 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.
Specific pcr products of 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. The clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products. The cloneez pcr cloning kit is designed for the quick cloning of pcr products without any headaches of restriction sites. The kit contains strataclone vector mix, strataclone cloning buffer. Mar 24, 2020 this method, sometimes using a different reporter gene, has been used to screen a variety of high fidelity pcr enzymes and to optimize pcr reaction conditions to minimize mutations 48. Molecular cloning produces identical, recombinant dna molecules for downstream applications, such as gene expression, protein purification, and mutational analysis. The clonejet pcr cloning kit is compatible with all pcr buffers supplied by thermo scientific. Molecular cloning using polymerase chain reaction, an. 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. Specific pcr products of pcr reaction mixture without any purification. Fiveminute cloning of taq polymeraseamplified pcr products.
Clonejet pcr cloning kit the thermo scientific clonejet pcr cloning kit is an advanced positiveselection system for highefficiency cloning of pcr products generated with any thermostable dna polymerase. Pcr cloning kit the clonejet pcr cloning kit is ideal for cloning blunt or stickyend pcr products. 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. Genscripts proprietary cloneez seamless cloning technology is a cuttingedge, recombinationbased cloning technology that can accurately and efficiently assemble genes and gene fragments in any combination of sequences. Clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna polymerase.
In vivo bluntend cloning through crisprcas9facilitated. Thermo scientific clonejet pcr cloning kit for 20 reactions. Using our proprietary cloneez enzyme, this kit is able to. The vector contains a lethal restriction enzyme gene. Sequence analysis, cloning and extracellular expression of. For sequence validation, the pcr product was subcloned using clonejet pcr cloning kit thermo scientific. 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. Feb 18, 2016 pcr and rt pcr products corresponding to mock or advtransduced myoblasts were purified by using the jetquick gel extraction spin kit genomed, according to the manufacturers recommendations and were ligated to the pjet1. In addition, the vector contains a t7 promoter for in vitro and in vivo transcription as well as sequencing of the insert. Purchase of this product conveys to the purchaser only the nontransferable right under these patents to use the product for research use only by the purchaser.
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