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10.1016/j.bbrep.2017.01.010 http://scihub22266oqcxt.onion/10.1016/j.bbrep.2017.01.010 C5614619!5614619!28956018     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Biochem+Biophys+Rep 2017 ; 9 (ä): 310-5 Nephropedia Template TP {{tp|p=28956018|t=2017. Evaluation of the efficiency and utility of recombinant enzyme-free seamless DNA cloning methods |pdf=|usr=}}{{28956018}} gab.com Text Evaluation of the efficiency and utility of recombinant enzyme-free seamless DNA cloning methods JO: Biochem Biophys Rep http://www.kidney.de/pget.php?&tw=1&pmid=28956018 #FOAvip Simple and low-cost recombinant enzyme-free seamless DNA cloning methods have recently become available. In vivo Escherichia coli cloning (iVEC) can directly transform a mixture of insert and vector DNA fragments into E. coli, which are ligated by endogenous homologous recombination activity in the cells. Seamless ligation cloning extract (SLiCE) cloning uses the endogenous recombination activity of E. coli cellular extracts in vitro to ligate insert and vector DNA fragments. An evaluation of the efficiency and utility of these methods is important in deciding the adoption of a seamless cloning method as a useful tool. In this study, both seamless cloning methods incorporated inserting DNA fragments into linearized DNA vectors through short (15?39 bp) end homology regions. However, colony formation was 30?60-fold higher with SLiCE cloning in end homology regions between 15 and 29 bp than with the iVEC method using DH5? competent cells. E. coli AQ3625 strains, which harbor a sbcA gene mutation that activates the RecE homologous recombination pathway, can be used to efficiently ligate insert and vector DNA fragments with short-end homology regions in vivo. Using AQ3625 competent cells in the iVEC method improved the rate of colony formation, but the efficiency and accuracy of SLiCE cloning were still higher. In addition, the efficiency of seamless cloning methods depends on the intrinsic competency of E. coli cells. The competency of chemically competent AQ3625 cells was lower than that of competent DH5? cells, in all cases of chemically competent cell preparations using the three different methods. Moreover, SLiCE cloning permits the use of both homemade and commercially available competent cells because it can use general E. coli recA? strains such as DH5? as host cells for transformation. Therefore, between the two methods, SLiCE cloning provides both higher efficiency and better utility than the iVEC method for seamless DNA plasmid engineering. Twit Text FOAVip Evaluation of the efficiency and utility of recombinant enzyme-free seamless DNA cloning methods ????Biochem Biophys Rep http://www.kidney.de/pget.php?&tw=1&pmid=28956018 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28956018 #FOAvip English Wikipedia <ref>{{Cite pmid|28956018}}</ref> Evaluation of the efficiency and utility of recombinant enzyme-free seamless DNA cloning methods #MMPMID28956018 Motohashi K Biochem Biophys Rep 2017[Mar]; 9 (ä): 310-5 PMID28956018show ga Simple and low-cost recombinant enzyme-free seamless DNA cloning methods have recently become available. In vivo Escherichia coli cloning (iVEC) can directly transform a mixture of insert and vector DNA fragments into E. coli, which are ligated by endogenous homologous recombination activity in the cells. Seamless ligation cloning extract (SLiCE) cloning uses the endogenous recombination activity of E. coli cellular extracts in vitro to ligate insert and vector DNA fragments. An evaluation of the efficiency and utility of these methods is important in deciding the adoption of a seamless cloning method as a useful tool. In this study, both seamless cloning methods incorporated inserting DNA fragments into linearized DNA vectors through short (15?39 bp) end homology regions. However, colony formation was 30?60-fold higher with SLiCE cloning in end homology regions between 15 and 29 bp than with the iVEC method using DH5? competent cells. E. coli AQ3625 strains, which harbor a sbcA gene mutation that activates the RecE homologous recombination pathway, can be used to efficiently ligate insert and vector DNA fragments with short-end homology regions in vivo. Using AQ3625 competent cells in the iVEC method improved the rate of colony formation, but the efficiency and accuracy of SLiCE cloning were still higher. In addition, the efficiency of seamless cloning methods depends on the intrinsic competency of E. coli cells. The competency of chemically competent AQ3625 cells was lower than that of competent DH5? cells, in all cases of chemically competent cell preparations using the three different methods. Moreover, SLiCE cloning permits the use of both homemade and commercially available competent cells because it can use general E. coli recA? strains such as DH5? as host cells for transformation. Therefore, between the two methods, SLiCE cloning provides both higher efficiency and better utility than the iVEC method for seamless DNA plasmid engineering. äEvaluation of the efficiency and utility of recombinant enzyme-free se(epmc).pdf DeepDyve Pubget Overpricing