Strataclone pcr cloning kit from stratagene biocompare. 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. The kit works well with both phosphorylated and nonphosphorylated dna fragments. The vector contains a lethal restriction enzyme gene that is disrupted by ligation. The clonejet pcr cloning kit contains a novel, readytouse positive selection cloning vector pjet1. Oct 09, 2006 the strataclone pcr cloning kit is designed for efficient cloning of pcr fragments. The cloneez pcr cloning kit is designed for the quick cloning of pcr products without any headaches of restriction sites.
The thermo scientific clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna. Clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any thermostable dna polymerase. The strataclone pcr cloning kit is designed for efficient cloning of pcr fragments. In vivo bluntend cloning through crisprcas9facilitated. According to kit self ligated vector should not grow. Yingguang frank chans lab contains the insert glikruppel family member gli3 and is published in unpublished this plasmid is available through addgene. Primers are also provided, allowing screening for inserts by colony pcr andor. Any other blunt or stickyend dna fragment can be cloned as well. Specific pcr products of 3 kb, ligation can be prolonged to 30 min. By using this kit, we can select specific pairs of restriction enzyme sites at both the 5 and 3 ends of the desired gene to clone into a vector of interest. Sequence analysis, cloning and extracellular expression of. The traditional path for orf cloning has usually started with experimental observations driving the identification of one or several genes of interest to a. Gelanalyze the pcr product for specificity and yield before cloning.
For qiagen pcr cloning kit qiagen pcr cloningplus kit pdf 771kb english format. 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. Any other blunt or stickyend dna fragment can be cloned. The clonejet pcr cloning kit is an advanced positive selection system for the highest. The clonejet pcr cloning kit is compatible with all pcr buffers supplied by thermo scientific. For convenience in mapping and manipulation of the insert, the pjet1. See attached the thermo scientific clonejet pcr cloning kit. Purified amplicons were then subcloned using the clonejet pcr cloning kit thermofisher scientific, and subsequently transformed into. 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. May 03, 2019 the pcr fragment was cloned subsequently using the clonejet pcr cloning kit fermentas.
Thermo scientific clonejet pcr cloning kit for 20 reactions. 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. Thermo scientific clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products generated with any. Pcr and rtpcr products corresponding to mock or advtransduced myoblasts were purified by using the jetquick gel extraction spin kit genomed, according to the. Pcr enzyme recommendations for specific applications are listed in table 1. The vector contains a lethal restriction enzyme gene. 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. This kit utilizes the 3adenosine overhangs produced during amplification. The vector contains a lethal restriction enzyme gene that is disrupted by ligation of a dna insert into the cloning site. 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.
For sequence validation, the pcr product was subcloned using clonejet pcr cloning kit thermo scientific. 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. 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. Specific pcr products of pcr reaction mixture without any purification. 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. It is ideal for phosphorylated or nonphosphorylated dna fragments. 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. 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.
Choochoo cloning tm kits are highly efficient directional pcr cloning kits for rapid, ligase and restriction enzymeindependent cloning of pcr products. An 8 l aliquot of the recovered pcr sample was ligated into pjet1. An aliquot of this reaction was separated and recovered using the flashgel recovery system. Using our proprietary cloneez enzyme, this kit is able to. 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. All components of the thermo scientific clonejet pcr cloning kit. The clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products.
The kit contains strataclone vector mix, strataclone cloning buffer, solopack pcr cloning competent cells, strataclone control insert and puc18 control plasmid. Clonejet pcr cloning kit k1231 from thermo fisher scientific. Clonejet pcr cloning kit with dh10b competent cells. The kit contains a positive selection vector ensuring that only bacterial cells. Sixty independent clones were analyzed by colony pcr using the smartiiaprimer and the genespecific gldpa race6 oligonucleotide 5. For qiagen pcr cloning kit qiagen pcr cloningplus kit pdf 771kb english format file size language download get adobe reader contact qiagen. Primers are also provided, allowing screening for inserts by colony pcr andor sequencing. Strataclone pcr cloning kit 5 map for the strataclone pcr cloning vector pscaampkan feature nucleotide position. Specific pcr products of may 01, 2019 the clonejet pcr cloning kit is an advanced positive selection system for highefficiency cloning of pcr products.
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. Pdf reproducibility of vibrionaceae population structure in. Jan 19, 2015 for sequence validation, the pcr product was subcloned using clonejet pcr cloning kit thermo scientific. Combine the following reagents sequentially on ice. 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. As a result, only bacterial cells with recombinant plasmids are able to form colonies. The high quality ta cloning vector ptz57rt is ready. The kit contains strataclone vector mix, strataclone cloning buffer. Pcr cloning kit the clonejet pcr cloning kit is ideal for cloning blunt or stickyend pcr products. Mar 30, 2010 pcr cloning procedure consisting of the four following steps. Molecular cloning using polymerase chain reaction, an.
Transformation analysis of recombinant clones the clonejet pcr c loning kit is compatible with all common li laboratory strains. When i cloned pcr products in pjet i plate something like 2550 l of the transformation and the plates are full with colonies. The pcr fragment was cloned subsequently using the clonejet pcr cloning kit fermentas. 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. 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. Pcr cloning procedure consisting of the four following steps.
Clonejet an advanced positive selection system for high. The cells are provided in a convenient singletube transformation format. 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. The clonejet pcr cloning kit is an advanced positive selection system. The high quality ta cloning vector ptz57rt is ready to use for efficient ligation with pcr products providing high cloning yields and low background. Ligation times longer than 30 min are not recommended and may decrease cloning efficiency. The neb pcr cloning kit contains a sufficient supply of materials to perform 20 x 10. Molecular cloning produces identical, recombinant dna molecules for downstream applications, such as gene expression, protein purification, and mutational analysis. Hi, i wanted to ask, how can i release an inserted dna into pjet1. The thermo scientific instaclone pcr cloning kit is a ta system for direct onestep cloning of pcr products with 3da overhangs 1. Fiveminute cloning of taq polymeraseamplified pcr products. 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. Followed thermo scientific clonejet pcr bluntend cloning protocol. 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.
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