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Methods of processing Oxford Nanopore sequencing data for metagenomics
Barilíková, Lujza ; Provazník, Ivo (referee) ; Kupková, Kristýna (advisor)
The revolutionary sequencing technology introduced by Oxford Nanopore Technologies – MinION holds a great promise in the field of metagenomics. Low cost, produced long reads and portability, due to its small dimensions, represents only one of the many advantages of this technology. Despite the benefits, there is a lack of available computational tools for handling the produced data. The theoretical part of the thesis first introduces current sequencing technologies with main focus on the third-generation sequencing and especially on nanopore sequencing. The recent possibilities of metagenomic data visualization are introduced. The main purpose of the bachelor thesis is to make an algorithm for binning of metagenomic samples based on use of dimensionality reduction techniques straight on raw data produced by nanopore sequencing.
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Direct Binning of Metagenomic Signals from Nanopore Sequencing
Lebó, Marko ; Jugas, Robin (referee) ; Sedlář, Karel (advisor)
This diploma thesis deals with taxonomy independent methods for classification of metagenomic signals, aquired by a MinION sequencer. It describes the formation and character of metagenomic data and already existing methods of metagenomic data classification and their development. This thesis also evaluates an impact of the third generation sequencing techniques in the world of metagenomics and further specialises on the function of the Oxford Nanopore MinION sequencing device. Lastly, a custom method for metagenomic data classification, based on data obtained from a MinION sequencer, is proposed and compared with an already existing method of classification.
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Next generation sequencing in clinical virology: method optimization and it's use for samples with unknown infectious agent
Poláčková, Kateřina ; Kramná, Lenka (advisor) ; Nunvář, Jaroslav (referee)
The use of the MinION sequencer (Oxford Nanopore) was tested on samples prepared to simulate infectious samples. The tested procedure is to simulate work with a sample with an unknown pathogen. Therefore, a metagenomic approach was chosen. Three kits were tested: Rapid Barcoding Sequencing, PCR Barcoding and Premium whole genome amplification. Each kit differed in duration, difficulty to prepare and in amplification of nucleic acids. In total it was chosen eight viruses with different genome lengths and with varying types of the genome (5,6 - 152 kb, ss/ds RNA, dsDNA). Ten samples were prepared to simulate different types of infection (respiratory, gastrointestinal tract and urine), and one sample contained pure water as a negative control. Before preparation of the library with Oxford Nanopore's kits, DNase/RNase treatment was used. The viral RNA was transcribed into DNA and in chosen samples were amplificated to reach a higher concentration of nucleic acids. Rapid barcoding sequencing kit detected all selected viruses with the highest number of viral reads (4403) with a length between 100 and 250 nt and quality coverage of viral genomes. PCR Barcoding kit detected five out of eight viruses, and the number of identified reads with a length of 100-200 nt distinctly decreased. Premium whole genome...
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Signal Based Processing Of Metagenomic Data From Nanopore Sequencing
Kupková, Kristýna
The revolutionary sequencing technology introduced by Oxford Nanopore Technologies – MinION holds a great promise in the field of metagenomics due to its low cost, produced long reads and small size, which makes it available also for field work. The only problem preventing this technology from reaching its full potential is the lack of available computational tools for handling the produced data. Here we present an algorithm for processing of the raw signal sequences generated by nanopore sequencing for metagenomic purposes allowing to remove viral sequences from a contaminated metagenomic sample.
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Direct Binning of Metagenomic Signals from Nanopore Sequencing
Lebó, Marko ; Jugas, Robin (referee) ; Sedlář, Karel (advisor)
This diploma thesis deals with taxonomy independent methods for classification of metagenomic signals, aquired by a MinION sequencer. It describes the formation and character of metagenomic data and already existing methods of metagenomic data classification and their development. This thesis also evaluates an impact of the third generation sequencing techniques in the world of metagenomics and further specialises on the function of the Oxford Nanopore MinION sequencing device. Lastly, a custom method for metagenomic data classification, based on data obtained from a MinION sequencer, is proposed and compared with an already existing method of classification.
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Methods Of Processing Oxford Nanopore Sequencing
Data For Metagenomics
Barilíková, Lujza
The presented paper describes a new method of processing data produced by revolutionary sequencing technology introduced by Oxford Nanopore Technologies – MinION, which holds a great promise in the field of metagenomics. Low cost, produced long reads and portability, due to its small dimensions, represents only one of the many advantages of this technology. Despite of the benefits, there is a lack of available computational tools for handling the produced data and that is the reason, why a new method of processing such data should be created. In this study such method is created based on dimensionality reduction for data visualization.
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Methods of processing Oxford Nanopore sequencing data for metagenomics
Barilíková, Lujza ; Provazník, Ivo (referee) ; Kupková, Kristýna (advisor)
The revolutionary sequencing technology introduced by Oxford Nanopore Technologies – MinION holds a great promise in the field of metagenomics. Low cost, produced long reads and portability, due to its small dimensions, represents only one of the many advantages of this technology. Despite the benefits, there is a lack of available computational tools for handling the produced data. The theoretical part of the thesis first introduces current sequencing technologies with main focus on the third-generation sequencing and especially on nanopore sequencing. The recent possibilities of metagenomic data visualization are introduced. The main purpose of the bachelor thesis is to make an algorithm for binning of metagenomic samples based on use of dimensionality reduction techniques straight on raw data produced by nanopore sequencing.
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