Hoinka Jan, Przytycka Teresa
National Center of Biotechnology Information, National Library of Medicine, NIH, Bethesda, MD 20894, USA.
National Center of Biotechnology Information, National Library of Medicine, NIH, Bethesda, MD 20894, USA.
Methods. 2016 Aug 15;106:82-5. doi: 10.1016/j.ymeth.2016.04.011. Epub 2016 Apr 11.
Aptamers, short and synthetic RNA/DNA molecules binding distinct targets with high affinity and specificity, are identified via Systematic Evolution of Ligands by Exponential Enrichment (SELEX), an in vitro procedure that, starting from a pool of random ssDNA/RNA sequences, selects sequences by amplifying target-affine species through a series of selection cycles. This versatile protocol has recently been combined with high throughput sequencing, allowing arbitrary stages of the selection to be sequenced and analyzed in silico. As a prerequisite, these data require extensive preprocessing by means of quality controls, error correction and demultiplexing, all while taking into account the specific design of aptamers. Existing solutions addressing this task are currently present only as integrated components in larger pipelines, limiting their applicability in independent software solutions. Here we present AptaPLEX, a standalone and platform independent demultiplexer specifically designed for HT-SELEX data. Given the multiplexed data from one or multiple HT-SELEX experiments, AptaPLEX extracts and restores aptamers into the original selection cycles by identifying the barcode and primer regions in each read. AptaPLEX is capable of fuzzy matching for both the barcode and primers, and automatically corrects mismatches between forward and reverse reads for paired-end data. Our software provides a rich set of additional features and can easily be integrated into existing analysis automation pipelines on multiple platforms ranging from desktop machines to cloud based solutions.
适体是短的合成RNA/DNA分子,能以高亲和力和特异性结合不同的靶标,通过指数富集配体系统进化技术(SELEX)来鉴定,这是一种体外方法,从随机单链DNA/RNA序列库开始,通过一系列选择循环扩增与靶标亲和的物种来选择序列。这个通用方案最近已与高通量测序相结合,使得选择的任意阶段都能进行测序并在计算机上进行分析。作为前提条件,这些数据需要通过质量控制、纠错和解复用进行广泛的预处理,同时还要考虑适体的具体设计。目前解决此任务的现有解决方案仅作为较大流程中的集成组件存在,限制了它们在独立软件解决方案中的适用性。在此,我们展示了AptaPLEX,这是一款专门为HT-SELEX数据设计的独立且与平台无关的解复用器。给定来自一个或多个HT-SELEX实验的多路复用数据,AptaPLEX通过识别每个读段中的条形码和引物区域,提取并将适体恢复到原始选择循环中。AptaPLEX能够对条形码和引物进行模糊匹配,并自动校正双端数据中正向和反向读段之间的错配。我们的软件提供了丰富的附加功能,并且可以轻松集成到从台式机到基于云的解决方案等多个平台上的现有分析自动化流程中。