测序质量分值用来评估碱基的错误检出率。利用边合成边测序(SBS)技术,read中的每个碱基会得到基于phred样算法1,2与最初为桑格测序实验开发的算法类似)的质量分值。
了解Illumina SBS如何工作。
指定碱基的测序质量分值Q是按照如下公式定义的:
Q = -10log10(e)
其中e是预估的碱基错误检出率。
如下所示,20的质量分值代表了错误率为1/100,对应的检出准确率为99%。
质量分值 | 碱基的错误检出率 | 推断的碱基检出精确度 |
---|---|---|
10 (Q10) | 1/10 | 90% |
20 (Q20) | 1/100 | 99% |
30 (Q30) | 1/1000 | 99.9% |
Illumina测序化学过程提供较高的精确度,绝大多数碱基得分在Q30及以上。这种精确度水平是多种测序应用的理想选择,包括临床研究。了解为何PhiX可以作为Illumina NGS运行中的质控品,进行运行质量监测。
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