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16: Bioinformatics

Last updated

Jan 3, 2021
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- 15.2: Protein Purification (Activity)
- 16.1: Introduction

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16.1: Introduction
Biological sequences are passed to software in a standardized format referred to as FASTA. FASTA is a plain text format that can be read in any text editor (TextEdit, Notepad, VIM, etc). Nucleic acids (DNA and RNA) and Proteins are represented by single-letter nucleotides (A, T, C, G) or single letter amino acid (20 amino acids). FASTA sequences begin with a > character in the first line and can contain multiple sequence entries all demarcated by a new line and a title line beginning with >.
16.2: Sequence Analysis
This page contains instructions on how to perform a sequence analysis by counting the ORFs using a FASTA file and the UGENE software.
16.3: In Silico Restriction
Restriction enzymes act as molecular scissors. The ones we use in Molecular Biology are those that cut within known sequences that occur often enough, yet rare enough to cut our DNA into analyzable fragments. Molecular Biologists often use 6-cutters. This means that the site of digestion is “restricted” to a recognition sequence of 6 nucleotides. These nucleotides are usually palindromic as discussed before.
16.4: In Silico PCR
Using the primer sequences, one can determine the size and/or location of a PCR product. This can be done using BLAST or with a program like UGENE. This page contains instructions on how to use BLAST and UGENE to determine the size and/or location of a PCR product.
16.5: Primer-BLAST
Primer-BLAST is a combination of a program called Primer3 that aids in the design of primers with specific properties and BLAST. Primer-BLAST allows for the construction of primers for qPCR where the user can specify the melting temperature, reduce the amount of self-priming, and span exon-exon junctions in order to avoid amplification of contaminating genomic DNA. This process ensures that the primers designed fall within your design parameters and most likely only amplify your interested gene.
16.6: Morphometic Analysis
Morphometrics (morpho– shape; metrics– measurements) is the use of physical measurements to determine the relatedness of organisms. With extinct organisms that have died out long ago, DNA extraction proves to be difficult. Likewise, prior to DNA technologies to analyze species, Linnean taxonomy was ascribed to organisms based on similarities in features.
16.7: Sequence Alignment and Tree Building
This page contains an informative video on how to perform a tree-building process on UGENE using MUSCLE and PhyML as well as the command line for the example file.

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