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10.1042/BSR20160608 http://scihub22266oqcxt.onion/10.1042/BSR20160608 C5333777!5333777 !28183872     free     free     free Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\28183872 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Biosci+Rep 2017 ; 37 (2 ): ä Nephropedia Template TP {{tp|p=28183872 |t=2017. Mega primer-mediated molecular cloning strategy for chimaeragenesis and long DNA fragment insertion |pdf=|usr=}}{{28183872 }} gab.com Text Mega primer-mediated molecular cloning strategy for chimaeragenesis and long DNA fragment insertion JO: Biosci Rep http://www.kidney.de/pget.php?&tw=1&pmid=28183872 #FOAvip Molecular cloning methods based on primer and overlap-extension PCR are widely used due to their simplicity, reliability, low cost and high efficiency. In this article, an efficient mega primer-mediated (MP) cloning strategy for chimaeragenesis and long DNA fragment insertion is presented. MP cloning is a seamless, restriction/ligation-independent method that requires only three steps: (i) the first PCR for mega primer generation; (ii) the second PCR for exponential amplification mediated by the mega primers and (iii) DpnI digestion and transformation. Most importantly, for chimaeragenesis, genes can be assembled and constructed into the plasmid vector in a single PCR step. By employing this strategy, we successfully inserted four DNA fragments (approximately 500 bp each) into the same vector simultaneously. In conclusion, the strategy proved to be a simple and efficient tool for seamless cloning. Twit Text FOAVip Mega primer-mediated molecular cloning strategy for chimaeragenesis and long DNA fragment insertion ????Biosci Rep http://www.kidney.de/pget.php?&tw=1&pmid=28183872 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28183872 #FOAvip English Wikipedia <ref>{{Cite pmid|28183872 }}</ref> Mega primer-mediated molecular cloning strategy for chimaeragenesis and long DNA fragment insertion #MMPMID28183872 Zhang H ; Liu CJ ; Jiang H ; Zhou L ; Li WY ; Zhu LY ; Wu L ; Meng E ; Zhang DY Biosci Rep 2017[Apr]; 37 (2 ): ä PMID28183872 show ga Molecular cloning methods based on primer and overlap-extension PCR are widely used due to their simplicity, reliability, low cost and high efficiency. In this article, an efficient mega primer-mediated (MP) cloning strategy for chimaeragenesis and long DNA fragment insertion is presented. MP cloning is a seamless, restriction/ligation-independent method that requires only three steps: (i) the first PCR for mega primer generation; (ii) the second PCR for exponential amplification mediated by the mega primers and (iii) DpnI digestion and transformation. Most importantly, for chimaeragenesis, genes can be assembled and constructed into the plasmid vector in a single PCR step. By employing this strategy, we successfully inserted four DNA fragments (approximately 500 bp each) into the same vector simultaneously. In conclusion, the strategy proved to be a simple and efficient tool for seamless cloning. |Cloning, Molecular/*methods [MESH] |DNA Primers/*genetics [MESH] |DNA/*genetics [MESH] |Genetic Vectors/genetics [MESH] |Mutagenesis, Insertional/genetics [MESH] |Polymerase Chain Reaction/methods [MESH]Mega primer-mediated molecular cloning strategy for chimaeragenesis an(epmc).pdf DeepDyve Pubget Overpricing