PT  - JOURNAL ARTICLE
AU  - Riccardo Ziraldo
AU  - Massa J. Shoura
AU  - Andrew Z. Fire
AU  - Stephen D. Levene
TI  - Deconvolution of Nucleic-acid Length Distributions: A Gel Electrophoresis Analysis Tool and Applications
AID  - 10.1101/636936
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 636936
4099  - http://biorxiv.org/content/early/2019/05/15/636936.short
4100  - http://biorxiv.org/content/early/2019/05/15/636936.full
AB  - Next-generation DNA-sequencing (NGS) technologies, which are designed to streamline the acquisition of massive amounts of sequencing data, are nonetheless dependent on various preparative steps to generate DNA fragments of required concentration, purity, and average size (molecular weight). Current automated electrophoresis systems for DNA- and RNA-sample quality control, such as Agilent’s Bioanalyzer ®and TapeStation ® products, are costly to acquire and use; they also provide limited information for samples having broad size distributions. Here we describe a software tool that helps determine the size distribution of DNA fragments in an NGS library, or other DNA sample, based on gel-electrophoretic line profiles. The software, developed as an ImageJ plug-in, allows for straightforward processing of gel images, including lane selection and fitting of univariate functions to intensity distributions. The user selects the option of fitting either discrete profiles in cases where discrete gel bands are visible, or continuous profiles, having multiple bands buried under a single broad peak. The method requires only modest imaging capabilities and is a cost-effective, rigorous alternative characterization method to augment existing techniques for library quality control.