Better quality score compression through sequence-based quality smoothing

Current NGS techniques are becoming exponentially cheaper. As a result, there is an exponential growth of genomic data unfortunately not followed by an exponential growth of storage, leading to the necessity of compression. Most of the entropy of NGS data lies in the quality values associated to each read. Those values are often more diversified than necessary. Because of that, many tools such as Quartz or GeneCodeq, try to change (smooth) quality scores in order to improve compressibility without altering the important information they carry for downstream analysis like SNP calling

Lossy Compression of Quality Values in Sequencing Data

The dropping cost of sequencing human DNA has allowed for fast development of several projects around the world generating huge amounts of DNA sequencing data. This deluge of data has run up against limited storage space, a problem that researchers are trying to solve through compression techniques. In this study we address the compression of SAM files, the standard output files for DNA alignment. We specifically study lossy compression techniques used for quality values reported in the SAM file and analyze the impact of such lossy techniques on the CRAM format. We present a series of experiments using a data set corresponding to individual NA12878 with three different fold coverages. We introduce a new lossy model, dynamic binning, and compare its performance to other lossy techniques, namely Illumina binning, LEON and QVZ. We analyze the compression ratio when using CRAM and also study the impact of the lossy techniques on SNP calling. Our results show that lossy techniques allow a better CRAM compression ratio. Furthermore, we show that SNP calling performance is not negatively affected and may even be boosted.Natural Sciences and Engineering Research Council of Canad

Compression of Structured High-Throughput Sequencing Data

Large biological datasets are being produced at a rapid pace and create substantial storage challenges, particularly in the domain of high-throughput sequencing (HTS). Most approaches currently used to store HTS data are either unable to quickly adapt to the requirements of new sequencing or analysis methods (because they do not support schema evolution), or fail to provide state of the art compression of the datasets. We have devised new approaches to store HTS data that support seamless data schema evolution and compress datasets substantially better than existing approaches. Building on these new approaches, we discuss and demonstrate how a multi-tier data organization can dramatically reduce the storage, computational and network burden of collecting, analyzing, and archiving large sequencing datasets. For instance, we show that spliced RNA-Seq alignments can be stored in less than 4% the size of a BAM file with perfect data fidelity. Compared to the previous compression state of the art, these methods reduce dataset size more than 40% when storing exome, gene expression or DNA methylation datasets. The approaches have been integrated in a comprehensive suite of software tools (http://goby.campagnelab.org) that support common analyses for a range of high-throughput sequencing assays.National Center for Research Resources (U.S.) (Grant UL1 RR024996)Leukemia & Lymphoma Society of America (Translational Research Program Grant LLS 6304-11)National Institute of Mental Health (U.S.) (R01 MH086883

Lossy Compressor preserving variant calling through Extended BWT

A standard format used for storing the output of high-throughput sequencing experiments is the FASTQ format. It comprises three main components: (i) headers, (ii) bases (nucleotide sequences), and (iii) quality scores. FASTQ files are widely used for variant calling, where sequencing data are mapped into a reference genome to discover variants that may be used for further analysis. There are many specialized compressors that exploit redundancy in FASTQ data with the focus only on either the bases or the quality scores components. In this paper we consider the novel problem of lossy compressing, in a reference-free way, FASTQ data by modifying both components at the same time, while preserving the important information of the original FASTQ. We introduce a general strategy, based on the Extended Burrows-Wheeler Transform (EBWT) and positional clustering, and we present implementations in both internal memory and external memory. Experimental results show that the lossy compression performed by our tool is able to achieve good compression while preserving information relating to variant calling more than the competitors. Availability: the software is freely available at https://github.com/veronicaguerrini/BFQzip.Comment: Proceedings of the 15th International Joint Conference on Biomedical Engineering Systems and Technologie

Compression of DNA sequencing data

With the release of the latest generations of sequencing machines, the cost of sequencing a whole human genome has dropped to less than US$1,000. The potential applications in several fields lead to the forecast that the amount of DNA sequencing data will soon surpass the volume of other types of data, such as video data. In this dissertation, we present novel data compression technologies with the aim of enhancing storage, transmission, and processing of DNA sequencing data. The first contribution in this dissertation is a method for the compression of aligned reads, i.e., read-out sequence fragments that have been aligned to a reference sequence. The method improves compression by implicitly assembling local parts of the underlying sequences. Compared to the state of the art, our method achieves the best trade-off between memory usage and compressed size. Our second contribution is a method for the quantization and compression of quality scores, i.e., values that quantify the error probability of each read-out base. Specifically, we propose two Bayesian models that are used to precisely control the quantization. With our method it is possible to compress the data down to 0.15 bit per quality score. Notably, we can recommend a particular parametrization for one of our models which—by removing noise from the data as a side effect—does not lead to any degradation in the distortion metric. This parametrization achieves an average rate of 0.45 bit per quality score. The third contribution is the first implementation of an entropy codec compliant to MPEG-G. We show that, compared to the state of the art, our method achieves the best compression ranks on average, and that adding our method to CRAM would be beneficial both in terms of achievable compression and speed. Finally, we provide an overview of the standardization landscape, and in particular of MPEG-G, in which our contributions have been integrated.Mit der Einführung der neuesten Generationen von Sequenziermaschinen sind die Kosten für die Sequenzierung eines menschlichen Genoms auf weniger als 1.000 US-Dollar gesunken. Es wird prognostiziert, dass die Menge der Sequenzierungsdaten bald diejenige anderer Datentypen, wie z.B. Videodaten, übersteigen wird. Daher werden in dieser Arbeit neue Datenkompressionsverfahren zur Verbesserung der Speicherung, Übertragung und Verarbeitung von Sequenzierungsdaten vorgestellt. Der erste Beitrag in dieser Arbeit ist eine Methode zur Komprimierung von alignierten Reads, d.h. ausgelesenen Sequenzfragmenten, die an eine Referenzsequenz angeglichen wurden. Die Methode verbessert die Komprimierung, indem sie die Reads nutzt, um implizit lokale Teile der zugrunde liegenden Sequenzen zu schätzen. Im Vergleich zum Stand der Technik erzielt die Methode das beste Ergebnis in einer gemeinsamen Betrachtung von Speichernutzung und erzielter Komprimierung. Der zweite Beitrag ist eine Methode zur Quantisierung und Komprimierung von Qualitätswerten, welche die Fehlerwahrscheinlichkeit jeder ausgelesenen Base quantifizieren. Konkret werden zwei Bayes’sche Modelle vorgeschlagen, mit denen die Quantisierung präzise gesteuert werden kann. Mit der vorgeschlagenen Methode können die Daten auf bis zu 0,15 Bit pro Qualitätswert komprimiert werden. Besonders hervorzuheben ist, dass eine bestimmte Parametrisierung für eines der Modelle empfohlen werden kann, die – durch die Entfernung von Rauschen aus den Daten als Nebeneffekt – zu keiner Verschlechterung der Verzerrungsmetrik führt. Mit dieser Parametrisierung wird eine durchschnittliche Rate von 0,45 Bit pro Qualitätswert erreicht. Der dritte Beitrag ist die erste Implementierung eines MPEG-G-konformen Entropie-Codecs. Es wird gezeigt, dass der vorgeschlagene Codec die durchschnittlich besten Kompressionswerte im Vergleich zum Stand der Technik erzielt und dass die Aufnahme des Codecs in CRAM sowohl hinsichtlich der erreichbaren Kompression als auch der Geschwindigkeit von Vorteil wäre. Abschließend wird ein Überblick über Standards zur Komprimierung von Sequenzierungsdaten gegeben. Insbesondere wird hier auf MPEG-G eingangen, da alle Beiträge dieser Arbeit in MPEG-G integriert wurden

Performance evaluation of lossy quality compression algorithms for RNA-seq data

Background Recent advancements in high-throughput sequencing technologies have generated an unprecedented amount of genomic data that must be stored, processed, and transmitted over the network for sharing. Lossy genomic data compression, especially of the base quality values of sequencing data, is emerging as an efficient way to handle this challenge due to its superior compression performance compared to lossless compression methods. Many lossy compression algorithms have been developed for and evaluated using DNA sequencing data. However, whether these algorithms can be used on RNA sequencing (RNA-seq) data remains unclear. Results In this study, we evaluated the impacts of lossy quality value compression on common RNA-seq data analysis pipelines including expression quantification, transcriptome assembly, and short variants detection using RNA-seq data from different species and sequencing platforms. Our study shows that lossy quality value compression could effectively improve RNA-seq data compression. In some cases, lossy algorithms achieved up to 1.2-3 times further reduction on the overall RNA-seq data size compared to existing lossless algorithms. However, lossy quality value compression could affect the results of some RNA-seq data processing pipelines, and hence its impacts to RNA-seq studies cannot be ignored in some cases. Pipelines using HISAT2 for alignment were most significantly affected by lossy quality value compression, while the effects of lossy compression on pipelines that do not depend on quality values, e.g., STAR-based expression quantification and transcriptome assembly pipelines, were not observed. Moreover, regardless of using either STAR or HISAT2 as the aligner, variant detection results were affected by lossy quality value compression, albeit to a lesser extent when STAR-based pipeline was used. Our results also show that the impacts of lossy quality value compression depend on the compression algorithms being used and the compression levels if the algorithm supports setting of multiple compression levels. Conclusions Lossy quality value compression can be incorporated into existing RNA-seq analysis pipelines to alleviate the data storage and transmission burdens. However, care should be taken on the selection of compression tools and levels based on the requirements of the downstream analysis pipelines to avoid introducing undesirable adverse effects on the analysis results. Document type: Articl

Lossy compression of quality scores in differential gene expression: A first assessment and impact analysis

High-throughput sequencing of RNA molecules has enabled the quantitative analysis of gene expression at the expense of storage space and processing power. To alleviate these prob- lems, lossy compression methods of the quality scores associated to RNA sequencing data have recently been proposed, and the evaluation of their impact on downstream analyses is gaining attention. In this context, this work presents a first assessment of the impact of lossily compressed quality scores in RNA sequencing data on the performance of some of the most recent tools used for differential gene expression

Compression algorithms for biomedical signals and nanopore sequencing data

The massive generation of biological digital information creates various computing challenges such as its storage and transmission. For example, biomedical signals, such as electroencephalograms (EEG), are recorded by multiple sensors over long periods of time, resulting in large volumes of data. Another example is genome DNA sequencing data, where the amount of data generated globally is seeing explosive growth, leading to increasing needs for processing, storage, and transmission resources. In this thesis we investigate the use of data compression techniques for this problem, in two different scenarios where computational efficiency is crucial. First we study the compression of multi-channel biomedical signals. We present a new lossless data compressor for multi-channel signals, GSC, which achieves compression performance similar to the state of the art, while being more computationally efficient than other available alternatives. The compressor uses two novel integer-based implementations of the predictive coding and expert advice schemes for multi-channel signals. We also develop a version of GSC optimized for EEG data. This version manages to significantly lower compression times while attaining similar compression performance for that specic type of signal. In a second scenario we study the compression of DNA sequencing data produced by nanopore sequencing technologies. We present two novel lossless compression algorithms specifically tailored to nanopore FASTQ files. ENANO is a reference-free compressor, which mainly focuses on the compression of quality scores. It achieves state of the art compression performance, while being fast and with low memory consumption when compared to other popular FASTQ compression tools. On the other hand, RENANO is a reference-based compressor, which improves on ENANO, by providing a more efficient base call sequence compression component. For RENANO two algorithms are introduced, corresponding to the following scenarios: a reference genome is available without cost to both the compressor and the decompressor; and the reference genome is available only on the compressor side, and a compacted version of the reference is included in the compressed le. Both algorithms of RENANO significantly improve the compression performance of ENANO, with similar compression times, and higher memory requirements.La generación masiva de información digital biológica da lugar a múltiples desafíos informáticos, como su almacenamiento y transmisión. Por ejemplo, las señales biomédicas, como los electroencefalogramas (EEG), son generadas por múltiples sensores registrando medidas en simultaneo durante largos períodos de tiempo, generando grandes volúmenes de datos. Otro ejemplo son los datos de secuenciación de ADN, en donde la cantidad de datos a nivel mundial esta creciendo de forma explosiva, lo que da lugar a una gran necesidad de recursos de procesamiento, almacenamiento y transmisión. En esta tesis investigamos como aplicar técnicas de compresión de datos para atacar este problema, en dos escenarios diferentes donde la eficiencia computacional juega un rol importante. Primero estudiamos la compresión de señales biomédicas multicanal. Comenzamos presentando un nuevo compresor de datos sin perdida para señales multicanal, GSC, que logra obtener niveles de compresión en el estado del arte y que al mismo tiempo es mas eficiente computacionalmente que otras alternativas disponibles. El compresor utiliza dos nuevas implementaciones de los esquemas de codificación predictiva y de asesoramiento de expertos para señales multicanal, basadas en aritmética de enteros. También presentamos una versión de GSC optimizada para datos de EEG. Esta versión logra reducir significativamente los tiempos de compresión, sin deteriorar significativamente los niveles de compresión para datos de EEG. En un segundo escenario estudiamos la compresión de datos de secuenciación de ADN generados por tecnologías de secuenciación por nanoporos. En este sentido, presentamos dos nuevos algoritmos de compresión sin perdida, específicamente diseñados para archivos FASTQ generados por tecnología de nanoporos. ENANO es un compresor libre de referencia, enfocado principalmente en la compresión de los valores de calidad de las bases. ENANO alcanza niveles de compresión en el estado del arte, siendo a la vez mas eficiente computacionalmente que otras herramientas populares de compresión de archivos FASTQ. Por otro lado, RENANO es un compresor basado en la utilización de una referencia, que mejora el rendimiento de ENANO, a partir de un nuevo esquema de compresión de las secuencias de bases. Presentamos dos variantes de RENANO, correspondientes a los siguientes escenarios: (i) se tiene a disposición un genoma de referencia, tanto del lado del compresor como del descompresor, y (ii) se tiene un genoma de referencia disponible solo del lado del compresor, y se incluye una versión compacta de la referencia en el archivo comprimido. Ambas variantes de RENANO mejoran significativamente los niveles compresión de ENANO, alcanzando tiempos de compresión similares y un mayor consumo de memoria

Novel methods for comparing and evaluating single and metagenomic assemblies

The current revolution in genomics has been made possible by software tools called genome assemblers, which stitch together DNA fragments “read” by sequencing machines into complete or nearly complete genome sequences. Despite decades of research in this field and the development of dozens of genome assemblers, assessing and comparing the quality of assembled genome sequences still heavily relies on the availability of independently determined standards, such as manually curated genome sequences, or independently produced mapping data. The focus of this work is to develop reference-free computational methods to accurately compare and evaluate genome assemblies. We introduce a reference-free likelihood-based measure of assembly quality which allows for an objective comparison of multiple assemblies generated from the same set of reads. We define the quality of a sequence produced by an assembler as the conditional probability of observing the sequenced reads from the assembled sequence. A key property of our metric is that the true genome sequence maximizes the score, unlike other commonly used metrics. Despite the unresolved challenges of single genome assembly, the decreasing costs of sequencing technology has led to a sharp increase in metagenomics projects over the past decade. These projects allow us to better understand the diversity and function of microbial communities found in the environment, including the ocean, Arctic regions, other living organisms, and the human body. We extend our likelihood-based framework and show that we can accurately compare assemblies of these complex bacterial communities. After an assembly has been produced, it is not an easy task determining what parts of the underlying genome are missing, what parts are mistakes, and what parts are due to experimental artifacts from the sequencing machine. Here we introduce VALET, the first reference-free pipeline that flags regions in metagenomic assemblies that are statistically inconsistent with the data generation process. VALET detects mis-assemblies in publicly available datasets and highlights the current shortcomings in available metagenomic assemblers. By providing the computational methods for researchers to accurately evalu- ate their assemblies, we decrease the chance of incorrect biological conclusions and misguided future studies