Team:Evry/Biology/GenomeAssembly

From 2014.igem.org

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<h1> De novo Genome assembly</h1>
<h1> De novo Genome assembly</h1>
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<h4> Assembly Strategy</h4>
<h4> Assembly Strategy</h4>
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Our DNA was sequenced by the Genoscope and the quality of the data analysed by the fastqc softvare, provide the link of the data
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In order to perform the genome assembly of our  bacteria we choose to used the velvet software. We choose 4 kmer size. For the first one at  31 we obtained  1493 contigs. for a kmer size of 65,  413 contigs are obtain. For the two last kmer tested, 97 and 113 we obtained 275 and 245 contigs.The prokka software is an automated tools for genome annotation. The run take less than ten minutes, as they was proposed on their articles.In order to control the quality of our DNA seq we choose to made first a Proteome comparisons with Pseudovibrio  FOBEG1 which is our reference strain. For that purpose we decided to look at our first genome version contains 275 contigs which was provide by the Velvet de novo assembly tools. For the 5456 proteins of Pseudovibrio FO-BEG1 5132 proteins  (which are our reference strain) are match with a prokka anotation CDS.
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5002 with a unique alignment 4936, 4687 with an alignement  > 90%, 4415  > 95% and 2525 > 99%. For that purpose we are sure that we sequence a strain related to the Pseudovibrio genus.
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After this proteome comparison, we try to improved our genome assembly by improved the kmer size and we see that the contigs numbers was reduced to 245. In  order to analyse the annotation and the quality of our genome annotation we look at 4 main specific annotation type such as antibiotic resitance, restriction enzyme, metals such as cadmium, copper and mercury, and other toxic compound such as phenol, nitrate, nitrite.
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Our stategy was to used the velvet software whi is a de novo assembler
 
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for that purpose I chose to to perform my analyse with the velvet software which is known to have good reference
 
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Revision as of 03:20, 18 October 2014

IGEM Evry 2014

Biology - Genome Assembly


De novo Genome assembly





Assembly Strategy



In order to perform the genome assembly of our bacteria we choose to used the velvet software. We choose 4 kmer size. For the first one at 31 we obtained 1493 contigs. for a kmer size of 65, 413 contigs are obtain. For the two last kmer tested, 97 and 113 we obtained 275 and 245 contigs.The prokka software is an automated tools for genome annotation. The run take less than ten minutes, as they was proposed on their articles.In order to control the quality of our DNA seq we choose to made first a Proteome comparisons with Pseudovibrio FOBEG1 which is our reference strain. For that purpose we decided to look at our first genome version contains 275 contigs which was provide by the Velvet de novo assembly tools. For the 5456 proteins of Pseudovibrio FO-BEG1 5132 proteins (which are our reference strain) are match with a prokka anotation CDS. 5002 with a unique alignment 4936, 4687 with an alignement > 90%, 4415 > 95% and 2525 > 99%. For that purpose we are sure that we sequence a strain related to the Pseudovibrio genus. After this proteome comparison, we try to improved our genome assembly by improved the kmer size and we see that the contigs numbers was reduced to 245. In order to analyse the annotation and the quality of our genome annotation we look at 4 main specific annotation type such as antibiotic resitance, restriction enzyme, metals such as cadmium, copper and mercury, and other toxic compound such as phenol, nitrate, nitrite.



Proteome Comparisons with FOBEG1



Antibiotic resistance





Restriction enzyme





Nitrate/nitrite





Cadmium





Copper





Mercuric





Phenol



Figure : Anaerobic pathway degradation for phenol



REFERENCE