caa a22 4500 378890484 CHVBK 20180305123453.0 cr unu---uuuuu 161128e20040405xx s 000 0 eng 10.1515/CCLM.2004.068 doi (NATIONALLICENCE)gruyter-10.1515/CCLM.2004.068 Generation of eight adjacent mutations in a single step using a site-directed mutagenesis kit [Elektronische Daten] [Anna Cabré, Josefa Girona, Guillermo Zalba, María-Ujué Moreno, Javier Díez, Lluís Masana] Studies that characterize transcriptional regulatory elements involve the site-directed mutagenesis methodology to generate deletions, insertions and point mutations. Commercial PCR-based system kits are widely used to introduce up to four base changes in a target sequence. Sequential mutagenesis experiments allow more than four bases to be altered. In this work, we show that the QuikChange site-directed mutagenesis kit developed by Stratagene, with an optimum primer design, can change eight adjacent bases. This approach allows us to study the effect of DNA sequence changes on functionality of specific sequences from gene target regions, including promoters, exons and introns. As a result of this methodology, a faster and cheaper way of introducing this number of mutations is achieved in a single step with only one pair of primers, thus reducing the possibility of potential random mutation in the rest of the target sequence. Copyright © 2004 by Walter de Gruyter GmbH & Co. KG Medical equipment & techniques nationallicence Medical diagnosis nationallicence Diseases & disorders nationallicence Cabré Anna aut Girona Josefa aut Zalba Guillermo aut Moreno María-Ujué aut Díez Javier aut Masana Lluís aut Clinical Chemistry and Laboratory Medicine Walter de Gruyter 42/4(2004-04-05), 384-386 1434-6621 42:4<384 2004 42 cclm https://doi.org/10.1515/CCLM.2004.068 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/CCLM.2004.068 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cabré Anna aut NATIONALLICENCE 700 1- Girona Josefa aut NATIONALLICENCE 700 1- Zalba Guillermo aut NATIONALLICENCE 700 1- Moreno María-Ujué aut NATIONALLICENCE 700 1- Díez Javier aut NATIONALLICENCE 700 1- Masana Lluís aut NATIONALLICENCE 773 0- Clinical Chemistry and Laboratory Medicine Walter de Gruyter 42/4(2004-04-05), 384-386 1434-6621 42:4<384 2004 42 cclm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter