![]() ![]() This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.įunding: This work was supported by the Swiss National Foundation grants 135754, 141898, 132815 and NCCR Structural Biology. Received: Accepted: SeptemPublished: October 30, 2013Ĭopyright: © 2013 Sun et al. Raghava, CSIR-Institute of Microbial Technology, India (2013) AAscan, PCRdesign and MutantChecker: A Suite of Programs for Primer Design and Sequence Analysis for High-Throughput Scanning Mutagenesis. Ĭitation: Sun D, Ostermaier MK, Heydenreich FM, Mayer D, Jaussi R, Standfuss J, et al. The software is distributed under GPLv3 licence and is available at. We hope that the open-source nature of our software and ready availability of freeware tools used for its development will facilitate its adaptation and further development. We have used AAscan software to design primers to make over 700 mutants, with a success rate of over 80%. We also describe additional software tools which are used to analyse a large number of sequencing results for the presence of desired mutations, as well as related software to design primers for ligation independent cloning. Here we present an open-source multi-platform software AAscan developed to design primers for this task according to a set of empirical rules such as melting temperature, overall length, length of overlap regions, and presence of GC clamps at the 3’ end, for any desired substitution. One of the time-consuming tasks encountered in application of this technique is the design of primers for site-directed mutagenesis. Scanning mutagenesis is a powerful protein engineering technique used to study protein structure-function relationship, map binding sites and design more stable proteins or proteins with altered properties.
0 Comments
Leave a Reply. |