Bio-Informatics step by step guide – How to perform multiple sequence alingment and phylogenetics analysis by using CLUSTAL-X, MEGA, GENEDOC software
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A multiple sequence alignment (MSA) is a sequence alignment of three or more biological sequences, generally protein, DNA, or RNA. In many cases, the input set of query sequences are assumed to have an evolutionary relationship by which they share a linkage and are descended from a common ancestor. From the resulting MSA, sequence homology can be inferred and phylogenetic analysis can be conducted to assess the sequences’ shared evolutionary origins. Visual depictions of the alignment as in the image at right illustrate mutation events such as point mutations (single amino acid or nucleotide changes) that appear as differing characters in a single alignment column, and insertion or deletion mutations (indels or gaps) that appear as hyphens in one or more of the sequences in the alignment. Multiple sequence alignment is often used to assess sequence conservation of protein domains, tertiary and secondary structures, and even individual amino acids or nucleotides.
Multiple sequence alignment also refers to the process of aligning such a sequence set. Because three or more sequences of biologically relevant length can be difficult and are almost always time-consuming to align by hand, computational algorithms are used to produce and analyze the alignments. MSAs require more sophisticated methodologies than pairwise alignment because they are more computationally complex. Most multiple sequence alignment programs use heuristic methods rather than global optimization because identifying the optimal alignment between more than a few sequences of moderate length is prohibitively computationally expensive.
Clustal is a series of widely used computer programs used in Bioinformatics for multiple sequence alignment.[2] There have been many versions of Clustal over the development of the algorithm that are listed below. The analysis of each tool and its algorithm are also detailed in their respective categories. Available operating systems listed in the sidebar are a combination of the software availability and may not be supported for every current version of the Clustal tools. Clustal Omega has the widest variety of operating systems out of all the Clustal tools.
Molecular Evolutionary Genetics Analysis (MEGA) is computer software for conducting statistical analysis of molecular evolution and for constructing phylogenetic trees. It includes many sophisticated methods and tools for phylogenomics and phylomedicine. It is licensed as proprietary freeware. The project for developing this software was initiated by the leadership of Masatoshi Nei in his laboratory at the Pennsylvania State University in collaboration with his graduate student Sudhir Kumar and postdoctoral fellow Koichiro Tamura.[1] Nei wrote a monograph (pp. 130) outlining the scope of the software and presenting new statistical methods that were included in MEGA. The entire set of computer programs was written by Kumar and Tamura. The personal computers then lacked the ability to send the monograph and software electronically, so they were delivered by postal mail. From the start, MEGA was intended to be easy-to-use and include solid statistical methods only.
MEGA version 2 (MEGA2), which was coauthored by an additional investigator Ingrid Jakobson, was released in 2001.[2] All the computer programs and the readme files of this version could be sent electronically due to advances in computer technology. Around this time, the leadership of the MEGA project was taken over by Kumar (now at Temple University) and Tamura (now at Tokyo Metropolitan University). The monograph Molecular Evolutionary Genetics Analysis was often used as a textbook for new ways to study molecular evolution.
MEGA has been updated and expanded several times and currently all these versions are available from the MEGA website. The latest release, MEGA7, has been optimized for use on 64-bit computing systems. MEGA is in two version. A graphical user interface is available as a native Microsoft Windows program. A command line version, MEGA-Computing Core (MEGA-CC), is available for native cross-platform operation. The method is widely used and cited. With millions of downloads across the releases, MEGA is cited in more than 85,000 papers. The 5th version has been cited over 25,000 times in 4 years
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very nice and excellent
Nice tutorial