Monday, 3 August 2009

DNA Sequence of our Phage


When we have our Phage we are ready to study it and to verify if this phage is suitable for Phage Therapy.

Outline


A good outline from HHMI is reported below with a small modification:


• extracting bacteriophages from a local soil sample or from
a fresch water sample or from a patient sample or from a sewage sample;


• purify bacteriophages by plating and growing on a lawn of bacteria;


• further purifying a single bacteriophage by picking plaques and making a concentrated stock;


• isolating the bacteriophage DNA;


• preparing the bacteriophage DNA for sequencing ;


• digesting a sample of bacteriophage DNA with restriction enzymes and analyzing the fragments by electrophoresis and comparing fragment pattern to other samples;


• preparing a bacteriophage sample for electron microscopy
;


• characterizing the bacteriophage type/structure from electron micrographs
;


• annotating segments of the genome to identify open reading frames, genes, and regulatory sequences;


• comparing this bacteriophage genome to other sequenced bacteriophage genomes;


• comparing and analyzing the amino acid sequence to reveal relationships between species;


• using Web-based tools and software programs for these activities;


• following up any unusual features that emerge, such as genes transferred from the host bacterial genome to the phage genome;




How do we Sequence DNA?


DNA Sequencing from Wikipedia


DNA Sequencing and Animations


The Sanger Method