– Shearing, ligate adapter
• Cluster generation
– Bridge PCR
• Sequencing on Genome Analyzer IIx
– RTA (Run Time Analysis)
• Analysis pipeline
– Offline analysis, alignment, SNPs calling, reads counting
Cluster generation: Bridge amplification
Bridge amplification cycle repeated till multiple bridges are formed
CONFIDENTIAL – DO NOT DISTRIBUTE
Cluster generation
Paired end sequencing
3’ ends are
blocked
forward strand is cleaved
CONFIDENTIAL – DO NOT DISTRIBUTE
Sequencing reverse strand
Hybridize sequencing
Perform sequencing on forward strand Re-generate reverse strand
Perform sequencing on reverse strand
CONFIDENTIAL – DO NOT DISTRIBUTE
Sample Prep - Resequencing
高通量测序技术及原理介绍
童贻刚 军事医学科学院 微生物流行病研究所 tong.yigang@