Dataset Information

FreeIbis: an efficient basecaller with calibrated quality scores for Illumina sequencers.

ABSTRACT: MOTIVATION: The conversion of the raw intensities obtained from next-generation sequencing platforms into nucleotide sequences with well-calibrated quality scores is a critical step in the generation of good sequence data. While recent model-based approaches can yield highly accurate calls, they require a substantial amount of processing time and/or computational resources. We previously introduced Ibis, a fast and accurate basecaller for the Illumina platform. We have continued active development of Ibis to take into account developments in the Illumina technology, as well as to make Ibis fully open source. RESULTS: We introduce here freeIbis, which offers significant improvements in sequence accuracy owing to the use of a novel multiclass support vector machine (SVM) algorithm. Sequence quality scores are now calibrated based on empirically observed scores, thus providing a high correlation to their respective error rates. These improvements result in downstream advantages including improved genotyping accuracy. AVAILABILITY AND IMPLEMENTATION: FreeIbis is freely available for use under the GPL (http://bioinf.eva.mpg.de/freeibis/). It requires a Python interpreter and a C++ compiler. Tailored versions of LIBOCAS and LIBLINEAR are distributed along with the package.

SUBMITTER: Renaud G

PROVIDER: S-EPMC3634191 | biostudies-other | 2013 May

REPOSITORIES: biostudies-other

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freeIbis: an efficient basecaller with calibrated quality scores for Illumina sequencers.

Renaud Gabriel G Kircher Martin M Stenzel Udo U Kelso Janet J

Bioinformatics (Oxford, England) 20130306 9

<h4>Motivation</h4>The conversion of the raw intensities obtained from next-generation sequencing platforms into nucleotide sequences with well-calibrated quality scores is a critical step in the generation of good sequence data. While recent model-based approaches can yield highly accurate calls, they require a substantial amount of processing time and/or computational resources. We previously introduced Ibis, a fast and accurate basecaller for the Illumina platform. We have continued active de ...[more]

PMID: 23471300

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Project description:BackgroundLow-coverage sequencing is a cost-effective way to obtain reads spanning an entire genome. However, read depth at each locus is low, making sequencing error difficult to separate from actual variation. Prior to variant calling, sequencer reads are aligned to a reference genome, with alignments stored in Sequence Alignment/Map (SAM) files. Each alignment has a mapping quality (MAPQ) score indicating the probability a read is incorrectly aligned. This study investigated the recalibration of probability estimates used to compute MAPQ scores for improving variant calling performance in single-sample, low-coverage settings.Materials and methodsSimulated tomato, hot pepper and rice genomes were implanted with known variants. From these, simulated paired-end reads were generated at low coverage and aligned to the original reference genomes. Features extracted from the SAM formatted alignment files for tomato were used to train machine learning models to detect incorrectly aligned reads and output estimates of the probability of misalignment for each read in all three data sets. MAPQ scores were then re-computed from these estimates. Next, the SAM files were updated with new MAPQ scores. Finally, Variant calling was performed on the original and recalibrated alignments and the results compared.ResultsIncorrectly aligned reads comprised only 0.16% of the reads in the training set. This severe class imbalance required special consideration for model training. The F1 score for detecting misaligned reads ranged from 0.76 to 0.82. The best performing model was used to compute new MAPQ scores. Single Nucleotide Polymorphism (SNP) detection was improved after mapping score recalibration. In rice, recall for called SNPs increased by 5.2%, while for tomato and pepper it increased by 3.1% and 1.5%, respectively. For all three data sets the precision of SNP calls ranged from 0.91 to 0.95, and was largely unchanged both before and after mapping score recalibration.ConclusionRecalibrating MAPQ scores delivers modest improvements in single-sample variant calling results. Some variant callers operate on multiple samples simultaneously. They exploit every sample's reads to compensate for the low read-depth of individual samples. This improves polymorphism detection and genotype inference. It may be that small improvements in single-sample settings translate to larger gains in a multi-sample experiment. A study to investigate this is ongoing.

Project description:BACKGROUND:Advanced sequencing machines dramatically speed up the generation of genomic data, which makes the demand of efficient compression of sequencing data extremely urgent and significant. As the most difficult part of the standard sequencing data format FASTQ, compression of the quality score has become a conundrum in the development of FASTQ compression. Existing lossless compressors of quality scores mainly utilize specific patterns generated by specific sequencer and complex context modeling techniques to solve the problem of low compression ratio. However, the main drawbacks of these compressors are the problem of weak robustness which means unstable or even unavailable results of sequencing files and the problem of slow compression speed. Meanwhile, some compressors attempt to construct a fine-grained index structure to solve the problem of slow random access decompression speed. However, they solve the problem at the sacrifice of compression speed and at the expense of large index files, which makes them inefficient and impractical. Therefore, an efficient lossless compressor of quality scores with strong robustness, high compression ratio, fast compression and random access decompression speed is urgently needed and of great significance. RESULTS:In this paper, based on the idea of maximizing the use of hardware resources, LCQS, a lossless compression tool specialized for quality scores, was proposed. It consists of four sequential processing steps: partitioning, indexing, packing and parallelizing. Experimental results reveal that LCQS outperforms all the other state-of-the-art compressors on all criteria except for the compression speed on the dataset SRR1284073. Furthermore, LCQS presents strong robustness on all the test datasets, with its acceleration ratios of compression speed increasing by up to 29.1x, its file size reducing by up to 28.78%, and its random access decompression speed increasing by up to 2.1x. Additionally, LCQS also exhibits strong scalability. That is, the compression speed increases almost linearly as the size of input dataset increases. CONCLUSION:The ability to handle all different kinds of quality scores and superiority in compression ratio and compression speed make LCQS a high-efficient and advanced lossless quality score compressor, along with its strength of fast random access decompression. Our tool LCQS can be downloaded from https://github.com/SCUT-CCNL/LCQSand freely available for non-commercial usage.

Dataset Information

FreeIbis: an efficient basecaller with calibrated quality scores for Illumina sequencers.

Publications

freeIbis: an efficient basecaller with calibrated quality scores for Illumina sequencers.

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