Illumina测序的原理和应用教案.pptx

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？;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;Hiseq2500;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 ;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!