Motivation: Capillary electrophoresis (CE) of nucleic acids is a workhorse technology underlying high-throughput genome analysis and large-scale chemical mapping for nucleic acid structural inference. Despite the wide availability of CE-based instruments, there remain challenges in leveraging their full power for quantitative analysis of RNA and DNA structure, thermodynamics, and kinetics. In particular, the slow rate and poor automation of available analysis tools have bottlenecked a new generation of studies involving hundreds of CE profiles per experiment. Results: We propose a computational method called high-throughput robust analysis for capillary electrophoresis (HiTRACE) to automate the key tasks in large-scale nucleic acid CE analysis, including the profile alignment that has heretofore been a rate-limiting step in the highest throughput experiments. We illustrate the application of HiTRACE on thirteen data sets representing 4 different RNAs, three chemical modification strategies, and up to 480 single mutant variants; the largest data sets each include 87,360 bands. By applying a series of robust dynamic programming algorithms, HiTRACE outperforms prior tools in terms of alignment and fitting quality, as assessed by measures including the correlation between quantified band intensities between replicate data sets. Furthermore, while the smallest of these data sets required 7 to 10 hours of manual intervention using prior approaches, HiTRACE quantitation of even the largest data sets herein was achieved in 3 to 12 minutes. The HiTRACE method therefore resolves a critical barrier to the efficient and accurate analysis of nucleic acid structure in experiments involving tens of thousands of electrophoretic bands.
Provide fast, accurate, and easy-to-use software for nucleic acid experimentalists entering a high-throughput era of structural analysis.
This project contains the HiTRACE software that allows users to accurately and automatically perform key quantitative analysis tasks involved in high-throughput capillary electrophoresis (CE) of nucleic acids. CE has become a workhorse technology underlying high-throughput experimental methods such as high-speed genome sequencing and large-scale footprinting for nucleic acid structural inference. Despite the wide availability of CE-based equipment, there remain challenges in leveraging the full power of CE for quantitative analysis of RNA and DNA structure. We developed HiTRACE in order to address this issue. See Preprint for more information.
The application is a collection of Matlab scripts, appropriate for reproducing the computational results presented in this paper. For best user experience, it is highly recommended to turn on the multithreading option supported by the Matlab environment, if a multicore machine is used. More detailed documentation, including a run-through with example data is here.See all Downloads