MOTIVATION: As nanopore technology reaches ever higher throughput and accuracy, it becomes an increasingly viable candidate for reading out DNA data storage. Nanopore sequencing offers considerable flexibility by allowing long reads, real-time signal analysis, and the ability to read both DNA and RNA. We need flexible and efficient designs that match nanopore's capabilities, but relatively few designs have been explored and many have significant inefficiency in read density, error rate, or compute time. To address these problems, we designed a new single-read per-strand decoder that achieves low byte error rates, offers high throughput, scales to long reads, and works well for both DNA and RNA molecules. We achieve these results through a novel soft decoding algorithm that can be effectively parallelized on a GPU. Our faster decoder allows us to study a wider range of system designs. RESULTS: We demonstrate our approach on HEDGES, a state-of-the-art DNA-constrained convolutional code. We implement one hard decoder that runs serially and two soft decoders that run on GPUs. Our evaluation for each decoder is applied to the same population of nanopore reads collected from a synthesized library of strands. These same strands are synthesized with a T7 promoter to enable RNA transcription and decoding. Our results show that the hard decoder has a byte error rate over 25%, while the prior state of the art soft decoder can achieve error rates of 2.25%. However, that design also suffers a low throughput of 183 s/read. Our new Alignment Matrix Trellis soft decoder improves throughput by 257× with the trade-off of a higher byte error rate of 3.52% compared to the state of the art. Furthermore, we use the faster speed of our algorithm to explore more design options. We show that read densities of 0.33 bits/base can be achieved, which is 4× larger than prior MSA-based decoders. We also compare RNA to DNA, and find that RNA has 85% as many error-free reads when compared to DNA. AVAILABILITY AND IMPLEMENTATION: Source code for our soft decoder and data used to generate figures is available publicly in the Github repository https://github.com/dna-storage/hedges-soft-decoder (10.5281/zenodo.11454877). All raw FAST5/FASTQ data are available at 10.5281/zenodo.11985454 and 10.5281/zenodo.12014515.

MOTIVATION: DNA-based data storage is a quickly growing field that hopes to harness the massive theoretical information density of DNA molecules to produce a competitive next-generation storage medium suitable for archival data. In recent years, many DNA-based storage system designs have been proposed. Given that no common infrastructure exists for simulating these storage systems, comparing many different designs along with many different error models is increasingly difficult. To address this challenge, we introduce FrameD, a simulation infrastructure for DNA storage systems that leverages the underlying modularity of DNA storage system designs to provide a framework to express different designs while being able to reuse common components. RESULTS: We demonstrate the utility of FrameD and the need for a common simulation platform using a case study. Our case study compares designs that utilize strand copies differently, some that align strand copies using multiple sequence alignment algorithms and others that do not. We found that the choice to include multiple sequence alignment in the pipeline is dependent on the error rate and the type of errors being injected and is not always beneficial. In addition to supporting a wide range of designs, FrameD provides the user with transparent parallelism to deal with a large number of reads from sequencing and the need for many fault injection iterations. We believe that FrameD fills a void in the tools publicly available to the DNA storage community by providing a modular and extensible framework with support for massive parallelism. As a result, it will help accelerate the design process of future DNA-based storage systems. AVAILABILITY AND IMPLEMENTATION: The source code for FrameD along with the data generated during the demonstration of FrameD is available in a public Github repository at https://github.com/dna-storage/framed, (https://dx.doi.org/10.5281/zenodo.7757762).