测序平台:Illumina
样本类型:胚胎脑部切片
公开日期:2022-08-01
更新日期:2022-08-01
项目编号 | 组织类型 | 物种 | 测序平台 | 文库设计 | 文件 |
MANUTEST | 脑 | 鼠 | Illumina | PE150 |
表1 测序数据统计
Type | Values |
Number of Reads | 356817224 |
Valid Barcodes | 301432357 |
Valid UMIs | 352248978 |
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。
表2 比对结果统计
Type | Values |
Reads Mapped to Genome | 92.03% |
Reads Mapped Confidently to Genome | 81.86% |
Reads Mapped Confidently to Intergenic Regions | 10.25% |
Reads Mapped Confidently to Intronic Regions | 3.3% |
Reads Mapped Confidently to Exonic Regions | 68.3% |
Reads Mapped Confidently to Transcriptome | 64.27% |
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。
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。
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 | 67.52% |
Number of Spots Under Tissue | 51.09% |
注:
Sequencing Saturation:测序饱和度;
Number of Spots Under Tissue:切片组织下Spots的个数。
表4 不同水平Spots统计
Level | 13 | 7 | 6 | 5 | 4 | 3 | 2 | 1 |
Number of SupSpots | 2607 | 9637 | 13449 | 20087 | 33106 | 64437 | 174072 | 721059 |
Median UMI Counts per SupSpot | 11199 | 3020 | 2162 | 1442 | 864 | 437 | 154 | 34 |
Median Genes per SupSpot | 4095 | 1631 | 1251 | 895 | 578 | 315 | 121 | 29 |
Total Genes Detected | 27561 | 27575 | 27583 | 27572 | 27577 | 27572 | 27575 | 27574 |
注:
Level:分辨率水平;
Number of SupSpots:一个或多个spot合并成的supspot个数;
Median UMI Counts per SupSpot:每个SupSpot的UMI中位数;
Median Genes per SupSpot:每个SupSpot中基因数目的中位数;
Total Genes Detected:基因总数。
百创S1000芯片可实现多级分辨率分析。通过调整supspot level参数,选取不同大小的spot点数作为分析单元,可以实现不同分辨率的数据分析和数据挖掘,最高分辨率supspot level 1可达到5um亚细胞水平。
表5 supspot level分辨率表
SupSpot Level | 分辨率 | Spot点数 |
13 | 100um | 469 |
7 | 50um | 127 |
6 | 42um | 91 |
5 | 35um | 61 |
4 | 27um | 37 |
3 | 20um | 19 |
2 | 10um | 7 |
1 | 5um | 1 |