Dual Indexを持つLibraryのシーケンサーセットアップ

1 2012 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, Sentrix, SeqMonitor, Solexa, TruSeq, 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. Dual index Library 2 index DNA library DNA pooling) de-multiplex (multiplex analysis) index DNA Single- index library 3 Dual- index Single- index ; 24 Dual- index .

2 96 MiSeq Output: max 8 Gbases : 1 Gbases : 1 Gbases : 1 Gbases 4 Dual- index P5 index 2 P7 index 1 DNA Dual- index library index 1 (i7): P7 8 12 index 2 (i5): P5 8 8 12 x 8 = 96 Single- index library 5 Nextera kit TruSeq Amplicon TruSeq HT sample prep kit TruSeq Amplicon Nextera kit (Nextera/TruSeq Amplicon) : 96well index1 12 index2 8 96 Illumina Dual- index index 1 index 2 6 Nextera kit TruSeq Amplicon TruSeq HT sample prep kit Illumina Dual- index TruSeq DNA Adapter Plate (DAP)

3 TruSeq HT kit Dual- index 96well 7 TruSeq HT Sample Prep Kits - TruSeq DNA HT sample prep kit Nextera Sample Prep Kits - Nextera DNA sample prep kit - Nextera XT sample prep kit - Nextera Exome sample prep kit TruSeq Amplicon Kits - TruSeq TSCA sample prep kit - TruSeq Amplicon cancer panel Dual index Illumina Dual- index 8 index Single- index Dual- index Single- index Dual- index 9 Region complementary to P5 grafting primer index 2 P5 primer DNA insert P7 primer index 1 P7 grafting primer Flow cell surface Dual- index 10 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Read 1 sequencing primer Flow cell surface Read1 index 2 index 1 Dual- index 11 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer Sequence P7 grafting primer Flow cell surface Read1 (Read1)

4 index 2 index 1 index 2 index 1 Read 1 sequencing primer Dual- index 12 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Sequence Flow cell surface Read1 index 2 index 1 Read 1 sequencing primer Dual- index 13 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Flow cell surface Index1 index 2 index 1 index 1 Sequencing Primer Dual- index 14 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer index 1 read 8 cycles Flow cell surface Index1 (Index1 read)

5 index 1 Sequencing Primer index 2 index 1 Dual- index 15 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Flow cell surface Index1 read index 2 index 1 Dual- index 16 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer index 2 Sequencing Primer P7 grafting primer Flow cell surface Index2 index 2 index 1 Dual- index 17 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer index 2 Sequencing Primer index 2 read 8 cycles P7 grafting primer Flow cell surface Index2 index 2 index 1 Dual- index 18 Dual- index 19 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Flow cell surface Read1 Sequencing Primer P5 grafting primer Read1 index 2 index 1 Dual- index 20 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer SBS Sequencing Primer Sequence Flow cell surface P5 grafting primer Read1 index 2 index 1 Dual- index 21 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer SBS Sequencing Primer Sequence Flow cell surface

6 P5 grafting primer Read1 index 2 index 1 Dual- index 22 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer index 1 Sequencing Primer Flow cell surface P5 grafting primer Index1 index 2 index 1 Dual- index 23 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Flow cell surface P5 grafting primer index 1 read 8 cycles Index1 index 2 index 1 index 1 Sequencing Primer Dual- index 24 Region complementary to P5 grafting primer P5 primer DNA insert P7 primer P7 grafting primer Flow cell surface P5 grafting primer index 1 read 8 cycles Index1 read index 2 index 1 Dual- index 25 Flow cell surface P5 grafting primer P5 grafting primer Dual- index 26 Flow cell surface 7 chemistry only cycles P5 grafting primer 7cycle Dual- index 27 7 chemistry only cycles index 2 - 8 cycles Flow cell surface P5 grafting primer Index2 DNA (Index2)

7 Dual- index 28 index 2 index read 8 cycles Flow cell surface P5 grafting primer PCR 7 chemistry only cycles Dual- index 29 Original strand New strand Flow cell surface DNA Dual- index 30 New strand Flow cell surface Read1 P5 Dual- index 31 Read 2 Sequencing Primer Flow cell surface Read2 Dual- index 32 Read 2 Sequencing Primer Sequence Flow cell surface Read2 Dual- index 33 index 1(i7) and index 2 (i5) Dual index 96 Dual- index (12 index 1 x 8 index 2) Dual- index TruSeq HT Nextera TruSeq Amplicon kit.

8 Dual index : Dual- index 34 HiSeq/Genome Analyzer 35 Dual- index Nextera sample prep kit TruSeq HT sample prep kit TruSeq Cluster Kit v2 GA TruSeq Cluster Kit v3 HS TruSeq SBS Kit v5 GA TruSeq SBS Kit v3 HS Dual- index Sequencing Primer Box Nextera TruSeq HT single flow cell cBot/cluster generation HiSeq/GA HiSeq/GA 36 TruSeq Dual- index Sequencing Primer Kit TruSeq Dual- index Sequencing Primer Kit for Single-Read flow cell HP10- Read1 sequencing primer mix HP12- index 1(i7) sequencing primer mix HP9- index 2 (i5)

9 Sequencing primer TruSeq Dual- index Sequencing Primer Kit for Paired-End flow cell HP10-Read1 sequencing primer mix HP12- index 1 (i7) sequencing primer mix HP11- Read2 sequencing primer mix FC-121-1003 PE-121-1003 Nextera Single-Read TruSeq HT flow cell Dual index TruSeq Nextera TruSeq Nextera 37 TruSeq HT Dual- index Sequencing Primer Kit, Single-read flow cell Nextera Dual- index Sequencing Primer Kit Single- Read FC R1: HP6/10 I1: HP8/12 I2: HP9 Paired-End Read FC R1: HP6/10 I1: HP8/12 I2: grafted P5 R2: HP7/11 Single- Read FC R1:HP10 I1: HP12 I2: HP9 Paired-End Read FC R1: HP10 I1: HP12 I2: grafted P5 R2: HP11 38 IEM SBS index read Single- index #19: HT2 #18: Diluted HP3- #17: HP8 HP12 Dual index #19: HT2 #18: Diluted HP3- 4mls #17: HP8 or HP12 #16: HP9 39 IEM SBS index read Single index #19: HT2 #18: Diluted HP3- #17: HP8 or HP12 Dual- index #19: HT2 #18: Diluted HP3- 4mls #17: HP8 HP12 #16.

10 HP9 40 IEM SBS index read Read1 index read sequencing Read2 Read2 SBS Read2 sequencing Single- index #19: HT2 #18: Diluted HP3 #17: HP8 HP12 Dual index #19: HT2 #18: Diluted HP3- 4mls #17: HP8 or HP12 #10: RMX 41 Single index #19: HT2 #18: Diluted HP3 #17: HP8 or HP12 Dual- index #19: HT2 #18: Diluted HP3- 4mls #17: HP8 HP12 #10: RMX IEM SBS index read Read1 index read sequencing Read2 Read2 SBS Read2 sequencing 42 IEM SBS index read Read1 index read sequencing Read2 Read2 SBS Read2 sequencing Single- index RMX Dual index Prepare PE reagents except RMX 43 IEM SBS index read Read1 index read sequencing Read2 Read2 SBS Read2 sequencing Single index Pr