自动化桑格分析管道(ASAP):一种以最小用户干扰快速分析桑格测序数据的工具。

Aditya Singh, P. Bhatia
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引用次数: 11

摘要

Sanger测序平台,如应用生物系统仪器,生成色谱文件。一般来说,对于一个序列的一个区域,我们同时使用正向和反向引物对该区域进行测序,这样我们就有两个序列需要比对,并在突变检测研究之前形成共识。这项工作是繁琐的,需要时间,特别是如果基因很大,有许多外显子。因此,我们设计了一个快速的自动化命令系统来过滤、构建和对齐一致序列,还可以选择提取外显子区域,在所有帧中翻译它们,并在很短的时间内从原始序列数据开始执行氨基酸比对。在自动化突变分析管道(ASAP)的全部功能中,它能够读取“*”。通过命令行界面,将ab1”色谱文件转换为FASTQ格式,对低质量区域进行裁剪,对反向序列进行反向补全,建立一致序列,使用参考外显子序列提取外显子区域,在所有帧中翻译序列,将核酸序列和氨基酸序列分别对齐为参考核酸序列和参考氨基酸序列。所有文件都被创建,并可用于进一步分析。ASAP将作为Python 3发布。可在https://github.com/aditya-88/ASAP执行。本文描述的版本为0.28。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Automated Sanger Analysis Pipeline (ASAP): A Tool for Rapidly Analyzing Sanger Sequencing Data with Minimum User Interference.
Sanger sequencing platforms, such as applied biosystems instruments, generate chromatogram files. Generally, for 1 region of a sequence, we use both forward and reverse primers to sequence that area, in that way, we have 2 sequences that need to be aligned and a consensus generated before mutation detection studies. This work is cumbersome and takes time, especially if the gene is large with many exons. Hence, we devised a rapid automated command system to filter, build, and align consensus sequences and also optionally extract exonic regions, translate them in all frames, and perform an amino acid alignment starting from raw sequence data within a very short time. In full capabilities of Automated Mutation Analysis Pipeline (ASAP), it is able to read "*.ab1" chromatogram files through command line interface, convert it to the FASTQ format, trim the low-quality regions, reverse-complement the reverse sequence, create a consensus sequence, extract the exonic regions using a reference exonic sequence, translate the sequence in all frames, and align the nucleic acid and amino acid sequences to reference nucleic acid and amino acid sequences, respectively. All files are created and can be used for further analysis. ASAP is available as Python 3.x executable at https://github.com/aditya-88/ASAP. The version described in this paper is 0.28.
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