Transcription of Sequencing By Synthesis (SBS)
1 2013 Illumina, Inc. All rights reserved. Illumina, IlluminaDx, BaseSpace, BeadArray, BeadXpress, cBot, CSPro, DASL, DesignStudio, Eco, GAIIx, Genetic Energy, Genome Analyzer, GenomeStudio, GoldenGate, HiScan, HiSeq, Infinium, iSelect, miseq , Nextera, NuPCR, SeqMonitor, Solexa, TruSeq, TruSight, VeraCode, the pumpkin orange color, and the Genetic Energy streaming bases design are trademarks or registered trademarks of Illumina, Inc. All other brands and names contained herein are the property of their respective owners. Sequencing By Synthesis (SBS) 2 DNA 1 DNA 96 DNA DNA SBS 30 DNA 3 DNA SBS 4 HiScanSQ GAIIx miseq HiSeq 1000/1500 HiSeq 2000/2500 Sequencing By Synthesis (SBS) 5 1.
2 DNA 2. DNA 3. 4. 6 1. DNA DNA DNA DNA Insert bp DNA P5, P7 DNA SP Index 7 DNA TruSeq DNA prep DNA (1 ug ) DNA bp 2 DNA 1. DNA 8 2013 7 Kit Details Nextera Nextera XT DNA 96 index DNA TruSeq DNA TruSeq nano TruSeq PCR-Free DNA 24(LT), 96(HT) TruSeq RNA (v2) TruSeq Stranded RNA mRNA 24 Stranded TruSeq Small RNA Small RNA, 48 Nextera Rapid Capture (Exome/Custom) Nextera Mate Pair TruSeq Amplicon (Custom/Cancer) 1.
3 DNA 9 2. DNA DNA DNA 10 1~8 (P5/P7) HiSeq HighOutput 2. DNA 11 2. DNA 12 1. DNA 2. PCR DNA 3.
4 1000 4. P7 2. DNA miseq DNA 13 Sequencing By Synthesis (SBS) 3 DNA 1 4 O PPP HN N O O 3 O DNA HN N O O 3 O 5 X OH 1 3. 14 5 C C C C C C G G G G T T T T T A A A A A C G C G T A T G C C G C A A T G T T 3 Cycle 2 Cycle 3, 4, Cycle 1 1 A A A A A G G G G G C C C C T T T C 3.
5 15 1 100um 20um miseq 1 1500 4. 16 FC 1- 4 = 28 clusters 4. 17 Quality scores (Q Scores) Q scores BaseCall Q Score Probability of error Q30 1 in 1000 Q20 1 in 100 1- 4 = T G C T 4. 18 T T G C A ASCII Character Q-scorePF (0,1)SequenceInstrumentRun IDLaneTileX-coordY-coordIndex #Read # DNA 4.
6 19 20 PE( ) 5 Read 1 (100bp) C T A A G A T C T T G C G 5 3 1 Read1 21 Read1 Single molecule array 22 Read2 C T A A G A T C T T G C G 5 5 3 1 R2 23 single read paired-end read 100bp 100bp 100bp 24 Index DNA DNA pooling) de-multiplex (multiplex analysis)
7 DNA 25 24 96 miseq Output: max 8 Gbases : 1 Gbases : 1 Gbases : 1 Gbases Index 26 Read 1 27 Read 1 Index Read 28 Read 1 Index Read Read 2 29 Dual Index Library pdf 30 DNA 1 2 3 7 8 9 4 5 6 T G C T A C G A T.
8 SBS 5 5 3 C T G A A C G A T C G T C T G A A C G A T C G T T C T G A A T (DNA/RNA) 31 / MyIllumina : Bentley et al. (2008) Accurate whole human genome Sequencing using reversible terminator chemistry. Nature 456: 53-59
