测序平台:Illumina
样本类型:小鼠睾丸切片
公开日期:2023/4/26
1、测序reads统计如下表:
表1 测序数据统计
Type | Values |
Number of Reads | 598,040,914 |
Valid Barcodes | 569,325,963 |
Valid UMIs | 592,672,836 |
注:
Number of Reads:reads总数;
Valid Barcodes:包含有效Barcode的reads数;
Valid UMIs:包含有效UMIs的reads数。
2、数据比对
表2 比对结果统计
Type | Values |
Reads Mapped to Genome | 97.25% |
Reads Mapped Confidently to Genome | 90.34% |
Reads Mapped Confidently to Intergenic Regions | 5.08% |
Reads Mapped Confidently to Intronic Regions | 4.92% |
Reads Mapped Confidently to Exonic Regions | 80.34% |
Reads Mapped Confidently to Transcriptome | 80.85% |
注:
Reads Mapped to Genomes:比对到参考基因组上的Reads在总Reads中占的比例;
Reads Mapped Confidently to Genome:比对到参考基因组并得到转录本GTF信息支持的Reads在总Reads中占的比例;
Reads Mapped Confidently to Intergenic Regions:比对到基因间区域的Reads在总Reads中占的比例;
Reads Mapped Confidently to Intronic Regions:比对到内含子区域的Reads在总Reads中占的比例;
Reads Mapped Confidently to Exonic Regions:比对到外显子区域的Reads在总Reads中占的比例;
Reads Mapped Confidently to Transcriptome:比对到已知参考转录本的Reads在总Reads中占的比例。
3、图像处理
每个玻片上都有Spots,实验时被组织切片覆盖,但是切片只会覆盖到部分Spots,实验时也只会获得覆盖区域下Spots中的基因表达。
4、Spots统计
空间转录组的基因表达定量,主要基于UMI计数来实现的。通过UMI可以区分一条read是属于生物学重复还是技术重复,能够有效地去除PCR效应。对每个Barcode下的基因去除重复的UMI,统计unique UMI数目即表示细胞基因的表达量。分析统计如下:
表3 Spots统计
Type | Values |
Sequencing Saturation | 50.55% |
Percent of Spots Under Tissue | 57.28% |
Fraction Reads in Spots Under Tissue | 73.70% |
注:
Sequencing Saturation:测序饱和度;
Percent of Spots Under Tissue:切片组织下Spots的比例;
Fraction Reads in Spots Under Tissue:切片组织下Reads的比例。
注:
Level:分辨率水平;
Number of SupSpots:一个或多个spot合并成的supspot个数;
Median Genes per SupSpot:每个SupSpot中基因数目的中位数;
Median UMI Counts per SupSpot:每个SupSpot的UMI中位数;
Total Genes Detected:基因总数。