技术简介
SeekOne® DD单细胞转录组+HS胞内菌测序基于微流控技术原理,利用Barcoded Beads实现单细胞的分离与标记,分别构建同一个样本的胞内菌16S rRNA文库和转录组文库。可同时捕获细胞的基因表达与胞内菌16S rRNA信息,获得不同细胞类型胞内菌的种类与组成差异,为进一步揭示细菌与细胞的互作研究提供了新手段。
技术简介
优势亮点
优化裂菌流程
提高细菌信号检出信噪比
一胞多检
揭示菌与宿主细胞互作特征图谱
巢式富集+单独建库
提高胞内菌信号检出灵敏度
技术流程

样本处理

油包水逆转录及标记

转录组cDNA扩增建库&16S富集建库

上机测序

数据分析和可视化

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技术应用
01
患者VS健康组细菌组成
患者VS健康组细菌分布
疾病诊断细菌标志物
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02
细菌对宿主表型的影响
细菌对宿主功能的影响
细菌对疾病微环境的影响
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03
治疗效果预测与评估
耐药预测与评估
新治疗策略开发
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数据表现
细胞数捕获1W左右,转录组测序100G左右,16S测序5G左右
Specles
Sample
Estmated Number Cells
Median Genes per Cell
Valid Barcodes
Sequencing Saturation
Fraction Reads in Cells
Specles:
Sample: 甲状腺癌
Estmated Number Cells: 11265
Median Genes per Cell: 1906
Valid Barcodes: 98.66%
Sequencing Saturation: 29.51%
Fraction Reads in Cells: 88.39%
Specles:
Sample: 甲状腺癌旁
Estmated Number Cells: 11230
Median Genes per Cell: 2531
Valid Barcodes: 98.52%
Sequencing Saturation: 36.65%
Fraction Reads in Cells: 87.92%
Specles:
Sample: 肠炎
Estmated Number Cells: 11211
Median Genes per Cell: 2219
Valid Barcodes: 96.86%
Sequencing Saturation: 21.76%
Fraction Reads in Cells: 84.00%
Specles:
Sample: 肠癌
Estmated Number Cells: 13755
Median Genes per Cell: 1888
Valid Barcodes: 96.67%
Sequencing Saturation: 35.18%
Fraction Reads in Cells: 80.84%
Specles:
Sample: 肠癌旁
Estmated Number Cells: 10257
Median Genes per Cell: 1662
Valid Barcodes: 97.72%
Sequencing Saturation: 39.96%
Fraction Reads in Cells: 65.00%
Specles
Sample
Number of Positive Cells
Valid Barcodes
Total UMIs Detected
TOP Bacteria
Ratio
Specles:
Specles:甲状腺癌
Specles:1427
Specles:97.53%
Specles:1923
Specles:葡萄球菌属、痤疮杆菌属、棒状杆菌属
Specles:~50%
Specles:
Specles:甲状腺癌旁
Specles:1915
Specles:96.86%
Specles:2687
Specles:痤疮杆菌属、葡萄球菌属、棒状杆菌属
Specles:~50%
Specles:
Specles:肠炎
Specles:6140
Specles:98.24%
Specles:15512
Specles:志贺氏菌属、小杆菌属、韦荣氏菌属
Specles:~80%
Specles:
Specles:肠癌
Specles:5749
Specles:96.83%
Specles:9589
Specles:副球菌属、痤疮丙酸杆菌属、不动杆菌属
Specles:~30%
Specles:
Specles:肠癌旁
Specles:2143
Specles:97.14%
Specles:2848
Specles:痤疮杆菌属、棒状杆菌属、草绿菌属
Specles:~50%
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