Efficient minimizer orders for large values of k using minimum decycling sets.

Minimizers are ubiquitously used in data structures and algorithms for efficient searching, mapping, and indexing of high-throughput DNA sequencing data. Minimizer schemes select a minimum k-mer in every L-long subsequence of the target sequence, where minimality is with respect to a predefined k-mer order. Commonly used minimizer orders select more k-mers than necessary and therefore provide limited improvement in runtime and memory usage of downstream analysis tasks. The recently introduced universal k-mer hitting sets produce minimizer orders with fewer selected k-mers. Generating compact universal k-mer hitting sets is currently infeasible for k > 13, and thus, they cannot help in the many applications that require minimizer orders for larger k Here, we close the gap of efficient minimizer orders for large values of k by introducing decycling-set-based minimizer orders: new minimizer orders based on minimum decycling sets. We show that in practice these new minimizer orders select a number of k-mers comparable to that of minimizer orders based on universal k-mer hitting sets and can also scale to a larger k Furthermore, we developed a method that computes the minimizers in a sequence on the fly without keeping the k-mers of a decycling set in memory. This enables the use of these minimizer orders for any value of k We expect the new orders to improve the runtime and memory usage of algorithms and data structures in high-throughput DNA sequencing analysis.

Efficient mapping of accurate long reads in minimizer space with mapquik.

DNA sequencing data continue to progress toward longer reads with increasingly lower sequencing error rates. We focus on the critical problem of mapping, or aligning, low-divergence sequences from long reads (e.g., Pacific Biosciences [PacBio] HiFi) to a reference genome, which poses challenges in terms of accuracy and computational resources when using cutting-edge read mapping approaches that are designed for all types of alignments. A natural idea would be to optimize efficiency with longer seeds to reduce the probability of extraneous matches; however, contiguous exact seeds quickly reach a sensitivity limit. We introduce mapquik, a novel strategy that creates accurate longer seeds by anchoring alignments through matches of k consecutively sampled minimizers (k-min-mers) and only indexing k-min-mers that occur once in the reference genome, thereby unlocking ultrafast mapping while retaining high sensitivity. We show that mapquik significantly accelerates the seeding and chaining steps-fundamental bottlenecks to read mapping-for both the human and maize genomes with [Formula: see text] sensitivity and near-perfect specificity. On the human genome, for both real and simulated reads, mapquik achieves a [Formula: see text] speedup over the state-of-the-art tool minimap2, and on the maize genome, mapquik achieves a [Formula: see text] speedup over minimap2, making mapquik the fastest mapper to date. These accelerations are enabled from not only minimizer-space seeding but also a novel heuristic [Formula: see text] pseudochaining algorithm, which improves upon the long-standing [Formula: see text] bound. Minimizer-space computation builds the foundation for achieving real-time analysis of long-read sequencing data.

Designing efficient randstrobes for sequence similarity analyses.

MOTIVATION: Substrings of length k, commonly referred to as k-mers, play a vital role in sequence analysis. However, k-mers are limited to exact matches between sequences leading to alternative constructs. We recently introduced a class of new constructs, strobemers, that can match across substitutions and smaller insertions and deletions. Randstrobes, the most sensitive strobemer proposed in Sahlin (Effective sequence similarity detection with strobemers. Genome Res 2021a;31:2080-94. https://doi.org/10.1101/gr.275648.121), has been used in several bioinformatics applications such as read classification, short-read mapping, and read overlap detection. Recently, we showed that the more pseudo-random the behavior of the construction (measured in entropy), the more efficient the seeds for sequence similarity analysis. The level of pseudo-randomness depends on the construction operators, but no study has investigated the efficacy. RESULTS: In this study, we introduce novel construction methods, including a Binary Search Tree-based approach that improves time complexity over previous methods. To our knowledge, we are also the first to address biases in construction and design three metrics for measuring bias. Our evaluation shows that our methods have favorable speed and sampling uniformity compared to existing approaches. Lastly, guided by our results, we change the seed construction in strobealign, a short-read mapper, and find that the results change substantially. We suggest combining the two results to improve strobealign's accuracy for the shortest reads in our evaluated datasets. Our evaluation highlights sampling biases that can occur and provides guidance on which operators to use when implementing randstrobes. AVAILABILITY AND IMPLEMENTATION: All methods and evaluation benchmarks are available in a public Github repository at https://github.com/Moein-Karami/RandStrobes. The scripts for running the strobealign analysis are found at https://github.com/NBISweden/strobealign-evaluation.

Minimizer-space de Bruijn graphs: Whole-genome assembly of long reads in minutes on a personal computer.

DNA sequencing data continue to progress toward longer reads with increasingly lower sequencing error rates. Here, we define an algorithmic approach, mdBG, that makes use of minimizer-space de Bruijn graphs to enable long-read genome assembly. mdBG achieves orders-of-magnitude improvement in both speed and memory usage over existing methods without compromising accuracy. A human genome is assembled in under 10 min using 8 cores and 10 GB RAM, and 60 Gbp of metagenome reads are assembled in 4 min using 1 GB RAM. In addition, we constructed a minimizer-space de Bruijn graph-based representation of 661,405 bacterial genomes, comprising 16 million nodes and 45 million edges, and successfully search it for anti-microbial resistance (AMR) genes in 12 min. We expect our advances to be essential to sequence analysis, given the rise of long-read sequencing in genomics, metagenomics, and pangenomics. Code for constructing mdBGs is freely available for download at https://github.com/ekimb/rust-mdbg/.

A Randomized Parallel Algorithm for Efficiently Finding Near-Optimal Universal Hitting Sets.

As the volume of next generation sequencing data increases, an urgent need for algorithms to efficiently process the data arises. Universal hitting sets (UHS) were recently introduced as an alternative to the central idea of minimizers in sequence analysis with the hopes that they could more efficiently address common tasks such as computing hash functions for read overlap, sparse suffix arrays, and Bloom filters. A UHS is a set of k-mers that hit every sequence of length L, and can thus serve as indices to L-long sequences. Unfortunately, methods for computing small UHSs are not yet practical for real-world sequencing instances due to their serial and deterministic nature, which leads to long runtimes and high memory demands when handling typical values of k (e.g. k>13). To address this bottleneck, we present two algorithmic innovations to significantly decrease runtime while keeping memory usage low: (i) we leverage advanced theoretical and architectural techniques to parallelize and decrease memory usage in calculating k-mer hitting numbers; and (ii) we build upon techniques from randomized Set Cover to select universal k-mers much faster. We implemented these innovations in PASHA, the first randomized parallel algorithm for generating nearoptimal UHSs, which newly handles k>13. We demonstrate empirically that PASHA produces sets only slightly larger than those of serial deterministic algorithms; moreover, the set size is provably guaranteed to be within a small constant factor of the optimal size. PASHA's runtime and memory-usage improvements are orders of magnitude faster than the current best algorithms. We expect our newly-practical construction of UHSs to be adopted in many high-throughput sequence analysis pipelines.