Illumina测序的原理和应用.pptVIP

Illumina 测序的原理和应用 Illumina测序流程 Adapter and Library 接头和文库 Illumina测序流程 什么是 flow cell？ Flow cell 即流动槽，如载玻片大小，有8个通道 即8条lane。 1. Single DNA libraries are hybridized to primer lawn and extended by polymerase 2. Double-stranded molecule is denatured and original template is washed away 3. New synthesized strand is covalently attached to flow cell surface to form a bridge and extended by polymerase 4. Double-strand bridge is formed 5. Double-strand bridge is denatured and form two copies of covalently bound single-stranded templates 6. Bridge amplification cycle repeat Multiple bridges are formed dsDNA bridges are denatured and reverse strands are cleaved 9. Cleaved reverse strands are washed away and leaving a cluster with forward strands only Illumina测序流程 10. Sequencing primer is hybridized to adapter sequence 11. Only one nucleotide is added every cycle as the 3’ end is blocked 12. Cleave the fluor and free the 3’ end then repeat above steps for next cycle 13. Sequenced strand is stripped off and the index primer is added for to seq index 3‘端 5‘端 Index primer 14. Repeat bridge amplification to form reverse strand after index sequencing 15. Sequencing the reverse strands 高通量测序的应用 高通量测序的应用 Thank you! 感谢聆听！ THANK YOU FOR WATCHING! 演示结束！