ABI公司测序仪的应用
, Ph.D.
Technical Product Specialist
chenyd@
提要
DNA测序仪应用概况
ABI遗传分析仪家族
1 4 16 48 96
测序仪三大功能
测序仪可以用来做什么？
Diagram from Celera Genomics
线粒体DNA测序
MITOMAP: A Human Mitochondrial Genome Database.
–Mouse Karyotype ZooWeb-Karyotypes home page Human Karyotype
医学测序
杂合子检测：发现新突变
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2006-11-17 |
© 2006 Applied Biosystems
细菌和真菌鉴定
MicroSeq® ID Analysis Software
MicroSeq® 500 16S Bacterial Identification Kit MicroSeq® D2 LSU Fungal Identification Kit
DNA Sequencing of the 16S ribosomal RNA gene Classification of Eubacteria, Yeasts,and Filamentous Fungi Sequence polymorphisms Bacterial category A biothreat agents Food pathogens Same day results
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2006-11-17 |
© 2006 Applied Biosystems
SAGE™ Analysis
基因组范围的基因表达图式分析 鉴定转录子(mRNA)
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2006-11-17 |
© 2006 Applied Biosystems
基于测序的HLA配型
Donor / Recipient compatibility prior to tissue transplantation HLA typing products - distributed directly by Celera Diagnostics for research purposes only.
1-866-235-3723
/cdx/HLA_Typing
Referenced to GENBANK: HLA–C 6p21.3 major histocompatibility complex, class I, C.
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2006-11-17 |
© 2006 Applied Biosystems
功能强大的片段分析
• 片段长度多态性
• AFLP和T-RFLP
• BAC指纹图谱 • 相对荧光定量
• LOH和QMPSF
• 构象分析扫描突变
• SSCP和CSCE
• InVivoScribe B细胞和T细胞clonality • TILLING
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2006-11-17 |
© 2006 Applied Biosystems
相对荧光定量
细胞淋巴瘤(cell lymphomas) 及其他肿瘤中的等位基因丢失 染色体不平衡 部分和全部染色体丢失
LOH (Loss of Heterozygosity) P53 17p13.1
非遗传的in-vivo LOH
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2006-11-17 |
© 2006 Applied Biosystems
寻找新的SNP位点
Mutation Profiling - SNP Discovery and Analysis
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2006-11-17 |
© 2006 Applied Biosystems
SNP扫描
SNPlex™ Genotyping System
SNaPShot® Multiplex System SNP Validation Linkage Mapping and Association Studies Pharmacogenomic Research
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2006-11-17 |
© 2006 Applied Biosystems
SSCP突变快速扫描
SSCP
Amplify region of interest Heat denature in HiDi formamide and NaOH
A T A T A T
G C G C
Rapidly chill and electrophoresis under non denaturing conditions
G
C
WT Wild Type
20
rfu
MUT
mobility
Mutant
© 2006 Applied Biosystems
2006-11-17 |
SSCP Workflow
Post PCR Purification
SSCP data on ABI 3130 Series
植物AFLP®分析
AFLP技术原理
AFLP工作流程
AB公司AFLP相关产品
AFLP Data Generated on AB Systems
Expt1:
Polymorphic peak
Common peak
Experiment 2: AFLP collaboration with U.S. customer
Customer Research Project Goals
Genotypes from AFLP Analysis of Hedysarum Lines
Hedysarum Variety Identification by AFLP Analysis
HO U5 01 09
HO U5 01 18
HO U5 01 11
HO U5 01 14
MicroSeq®细菌鉴定
500 bp试剂盒
全长基因试剂盒
FDA Draft Guidelines
分子生物学手段鉴定微生物
MicroSeq®Microbial Identification Kit
试剂盒组成
细菌鉴定策略
forward
reverse
MicroSeq®工作流程or
样品制备特别简单
MicroSeq®真菌鉴定真菌鉴定试剂盒
rDNA在真菌染色体上的排列
测序目标是D2
强项：MicroSeq®ID 软件
methylSEQr Bisulfite Conversion Kit 甲基化分析
重亚硫酸钠转化试剂盒
Applied Biosystems Products for Each Step of the Workflow
methylSEQr™Bisulfite Conversion Kit。