1. Error: could not find path to FAST5 for read xxxx
报错的具体格式:
Error: could not find path to FAST5 for read 0000145b-a8c3-43c0-95c9-fc4a1986a2a2
Please make sure that this path is accessible: start_time=2020-10-06T08:16:56Z
这个报错的原因是找不到对应read的fast5文件,解决方案有两种:
- 1.检查目录下是否存在fast5文件
- 2.检查nanopolish index运行后,fastq文件路径下是否生成了相关索引文件,如果没有生成,那么基本就是软件问题,请重新安装(conda安装的nanopolish可能会出现这种报错)。
2. The fast5 file is compressed with VBZ but the required plugin is not loaded
报错的具体格式:
The fast5 file is compressed with VBZ but the required plugin is not loaded. Please read the instructions here: https://github.com/nanoporetech/vbz_compression/issues/5
这个报错的原因是因为你试图用Nanopolish处理FAST5文件,但后者已经使用VBZ压缩算法进行了压缩,而在你当前的环境中没有加载或提供处理这种压缩的必要插件,解决方案如下:
- 先在下面的网址选择下载合适版本的VBZ软件:
https://github.com/nanoporetech/vbz_compression/releases
比如:
wget https://github.com/nanoporetech/vbz_compression/releases/download/1.0.2/ont-vbz-hdf-plugin_1.0.2-1.bionic_amd64.deb
- 解压 .deb 文件:
ar x ont-vbz-hdf-plugin_1.0.2-1.bionic_amd64.deb
- 提取文件,名称大致是data.tar.gz:
tar xf data.tar.gz
- 将文件导入搜索路径,根据实际情况修改
export HDF5_PLUGIN_PATH=path_to_your_dir_/hdf-plugin/usr/local/hdf5/lib/plugin/
3. "NNNNN" model kmer and "Inf" in nanopolish events output
这个其实不算报错,只是结果文件中出现Inf值和NNNNN的model kmer:
高亮的部分作者的回复:
We have a state in our model that allows us to ignore events that may be sequencing artifacts. When this state is used, we output NNNNNN in the model_kmer column. We recommend these events are ignored in any subsequent analysis.
翻译如下:我们的模型中有一个状态,允许我们忽略可能是测序误差的事件。当使用此状态时,我们在 model_kmer 列中输出 NNNNNN。我们建议在后续的分析中忽略这些事件。
使用grep删除这些事件:
grep -v 'inf' input_file.txt > output_file.txt
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