TruSeq Custom Amplicon Library Preparation Guide Part
Contents
1. gt 15 u Ilumina FC 130 1006 Procedure 1 ss 14 80 40 HYP HYP2. Illumina zu D 28 lh TruSeq Custom Amplicon Library Preparation Guide TruSeq V UL Revision History 15027983 2013 8 H
3. RV HELT IAP TruSeq Custom Amplicon Library Preparation Guide TruSeq CA d r Od UOeoIIdmV yod H D Protocol d FPU MIDI FPU FPU 7 Microseal AT 1IAP
4. TruSeqW Best Practices EUC TruSeqW Documentation amp Literature Ilumina TruSeq
5. ym a N 3 HT1 PAL MiSeq v2 594 ul 6 ul MiSeq v3 580 ul 20 ul Diluted Amplicon Library DAL Pooled Amplicon Library PAL PAL 15 25C Diluted Amplicon Library DAL MiSeq Diluted Amplicon Library DAL DAL 96 2 DAL 1 2
6. Application Specific IP Ilumina Illumina umina Illumina Ilumina Ilumina Illumina Ine Numina lumina Numina
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8. 2013 Illumina Inc All rights reserved Ilumina 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 Genetic Energy Illumina Inc For Research Use Only C Ilumina Inc Promega Corporation
9. 12 Removal of Unbound Oligos Library Normalization Wash 1 LNW1 29 Library Normalization Illumina Experiment Manager 54 MiSeq Sample Sheet Preparation MiSeq 56 Ilumina Amplicon Viewer TSCA 43 TruSeq Custom Amplicon Library Preparation Process Overview TruSeq FFPE DNA
10. E TruSeq Custom Amplicon Library Preparation Guide TruSeq E b 30 Zi e H co Od 1 Ge9IO Ad Protocol 28 13 14 15 16 17 18 19 c CLP 2 a 80 200 ul b 30 c d P20
11. 4 0 15027983 CJPN How Does the TruSeq Custom Amplicon Library Preparation Assay Work TruSeqW 1 DNA 96 DNA DNA DCH MiSeq Weeer t eegener 2 eem em em em em m A VEER GEN A 2 anaee P7 1 3 2 P5 P7
12. Complete Amplicon Sequencing Solution for MiSeq MiSeq DNA zi g C 15027983 C JPN DNA Input Recommendations DNA Fy Zy MBI T NEIE Gr DNA 150 bp 175 DNA bp 250 bp 425 bp FFPE DNA 150 bp 175 bp 15y 50 ng 250 ng 250 ng Input DNA Quantitation DNA Illumina PicoGreen DNA dsDNA UV gDNA
13. L TruSeq IAP i7 4 ul TruSeq 15027983 C JPN 12 96 TruSeq DNA Polymerase 1 TDP1 56 ul PCR Master Mix 2 PMM2 2 8 ml 1 PMM2 TDP1
14. v NW 2 1 19 Microseal B SGP 1 000 xg 1 Ale an D SE DNA Bioanalyzer SE qPCR EI qPCR Eur SAFE STOPPING POINT Sequencing Library qPCR Quantification Guide Library Pooling and MiSeq Sample Loading AM75eg MiSeq SGP 15 250 TruSeq
15. Estimated Time 20 20 Consumables ELM3 Extension Ligation Mix 3 1 15 25C Illumina f SW1 Stringent Water 1 1 Famy 2 RC Illumina UB1 Universal Buffer 1 1 2 8C Illumina SE VR Illumina 1 Illumina MIDI 1 15027983 C JPN Preparation 1 15 25 Extension Ligation Mix 4 ELM4 Extension Ligation Mix 4 ELM4 20 D 2 2 8 Stringent Water 1 SW1 Universal Buffer 1 UB1
16. Illumina a Ilumina G GE Ilumina Illumina b Illumina 3 Illumina 90 SEN E ESCH I Illumina
17. 48 TruSeq Custom Amplicon Library Preparation Kit Contents TruSeqW Download Your Manifest File MiSeq TruSeq MiSeq Mylllumina Custom Products
18. i 0 15027983 CJPN a b N 6 8 HYP HYP 4 Additional Resources 20C 1 000 xs 1 7 HYP Oligo Hybridization for Sequencing Reagent 2 OHS2 35 3 5 3 Oligo Hybridization for Sequencing Reagent 2 OHS2 Custom Amplicon Oligo Tube CAT Oligo Hybridization for Seq
19. TruSeq Custom Amplicon Library Preparation Guide TruSeq D 3 quowudmby CH Supporting Information MiSeq Sample Sheet Preparation MiSeq 94 MiSeq MiSeq Sample Sheet Quick Reference Guide MiSeg Illumina Illumina Experiment Manager MiSeq Targeted Resequencing TruSeq Amplicon Custom Amplicon Oligo Tube CAT Experienced User Card Lab Tracking Form DCH
20. Ilumina Ilumina IIlumina
21. E ZS TruSeq Custom Amplicon Library Preparation Guide TruSeq 111 On Ilumina Ilumina
22. 2 FPU 3 PPU MIDI 370 45 2 16 15027983 KETC JPN 4 FPU Indexed Amplification Plate TruSeq Custom Amplicon Library Preparation Guide TruSeq IAP 17 soSIJO punog JO UOI1B8TT UOISU9 X 1 A1 IR eier lo rn E A Protocol PCR Amplification PCR DNA PCR
23. Optional Bioanalyzer System 1 5 ml Sg J EL Agilent Technologies m Ey 9 F E H o o o di ECH ai a A J H Thermal Cycler Humina TruSeq 92 15027983 C JPN BIO RAD DNA Engine Tetrad 2 MJ Research DNA Engine Tetrad Eppendorf Mastercycler Pro S S 100C
24. Illumina Amplicon Viewer Amplicon Viewer Myllumina http www ilumina com help amplicon_viewer default htm 44 15027983 CJPN TruSeq Custom Amplicon Library Preparation Process Overview TruSeq LO Sen TruSeq Custom Amplicon Library Preparation Guide TruSeq Supporting Information Acronyms 4 TruSeq ACD1 Amplicon Control DNA 1 CAT Custom Amplicon Oligo Tube DAL Diluted Amplicon Library
25. DesignStudio MyIllumina Se Products DesignStudio for TruSeq Custom Amplicon Place Your Order DesignStudio MyIllumina TruSeq Custom Amplicon Calculator Illumina Web TruSeq Mylllumina MiSeq TSCA
26. DAL 5 15027983 C JPN iSeq Diluted Amplicon Library DAL MiSeq 14 Diluted Amplicon Library DAL MiSeq Load Samples 15 MiSeqg System User Guide Miseqg 7 f Ilumina CAT TruSeq Custom Amplicon Library Preparation Guide TruSeq LL tO POSIN dE ch GLL L SUIPEOT ole Doc Un pue suooq AJ6JdT1 98 15027983 C JPN Supporting Information GI VS
27. 8 20 1 000 xg 1 9 IAP 10 KORO PCR X PCR PCR CAT DNA DNA ipy ACD1 ACP1 Custom Amplicon Oligo Tube CAT 1 100 250 ng 150 175 bp 250 bp 425 bp DNA FFPE PCR X 2 50 99 ng DNA lt 96 9 d 15027983 CJPN 150 175 bp 250 bp 425 bp 97 384 29 385 768 28 769 1 536
28. Ilumina lumina Illumina 2 Ilumina Ilumina Core IP Ilumina A a Illumina Application Specific IP
29. Illumina CIZ SR dt uc ACD1 ACP1 Ilumina 9 ACP1 DNA Procedure 1 HYP HYbridization Plate 96 PCR ET HYP 1 DNA ACD1 5u TE TruSeq Custom Amplicon Library Preparation Guide TruSeq 0 GG Lena keete ED E OO OSHO JO UOIJeZIDMdAH Protocol 3 HYP
30. 2 P20 Library Normalization Wash 1 LNW1 DL a e E OD Oo OC y S l P20 Library Normalization Wash 1 LNW1 LNP 0 1N NaOH 1 30 Microseal B LNP LNP 1 800 rpm 5 5
31. D 15027983 CJPN Trasig Workflow Cru Sea TruSeq SAFE STOPPING POINT 1 TruSeqW PCR 15 Ee 20 ET 80 30 SS o EBT AR AMPure XP ACP1 BON K SEH LNP 1 30 20 e E Se LNA1 S i LNB1 LNW1 PIEJ LNS2 FPU NaOH SGP T MiSeq PF 45 10 BES EL
32. Gii Illumina d sE PS Gv Ilumina Ilumina G v c EN Illumina
33. Ilumina Humina Ilumina Illumina Ilumina IHlumina
34. Ilumina 3 CLP CLean up Plate MIDI D 15027983 C JPN 4 CLP AMPure XP Beads ACD1 ACP1 Custom Amplicon Oligo Tube CAT HACH 150 bp 175 bp 250 bp 425 bp 5 60 AMPure XP Beads ul 60 ul 60 ul 45 ul SD ol CLP HLT IAP PCR H 2 E LE i 4 Additional Resources
35. Normalization Beads 1 LNB1 Om RS Library Normalization Additives 1 LNA1 Library ormalization Beads 1 LNB1 Library Normalization Beads 1 LNB1 es Library ormalization Additives 1 LNA1 4 LNP LNA1 LNB1 45 U 5 Microseal B LNP 6 LNP 1 800 rpm 30 4
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37. 10 Stringent Water 1 SW1 6 a Stringent Water 1 SW1 45 yl amp b FPU 2 400 xg 2 2 Stringent Water 1 SW1 10 Stringent Water 1 SW1 7 MDI
38. E sisch E T 90 Si Illumina 15027983 C JPN 9 a Illumina Illumina SE E 9 9 d TIlumina G EE
39. Library Normalization Wash 1 LNW1 45 1 Microseal B LNP LNP 1 800 rpm 5 2 LNP Library Normalization Wash 1 LNW1 a Library Normalization Wash 1 LNW1 45 U Microseal B LNP LNP 1 800 rpm 5
40. Downloads Vi TruSeq Documentation amp Literature 15027983 C JPN Protocol introduction IEE EE 6 TruSeq WorkHow TruSeq A UE e LE EE 7 Hybridization of Oligo Pool 8 Removal of Unbound Oligos LLL LLL LLL L22 12 Extension Ligation of Bound Oligos 16 DCH Amplification DCH 18 PCR Clean Up PCR llllalaaaaaaaaan anana anaa nannaa aLaaa Aaaa Aaaa anaana anaana anaana anaana 24 Library Normalization 29 Library Pooling and MiSeq Sample Loading MiSeq e 34 A Si fl Seenen TruSeq Custom Amplicon Library Preparation Guide TruSeq
41. 1 2 3 Normalization Beads 1 LNB1 Procedure 1 90 96 Library Norma P1000 1 000 ul RTOC E MZ 3 n Libra ET L P1000 DEEN ry Normalization Beads 1 LNB1 n Additives 1 LNA1 CC Library H 15 20 Library Normalization Beads 1 LNB1 ization Additives 1 LNA1 4 4 ml 15 m on Additives
42. 1 Illumina Illumina A SU IN SE O E lumin b Imlmmina Ilumina Cer G
43. OHS2 ELM4 LNS2 DNA 3 DNA Input Recommendations DNA OHS2 100 ng DNA PCR 18 PCR Amplification PCR 34 Library Pooling and MiSeq Sample Loading MiSeq MiSeq v3 DAL 15027983 C JPN 15027983 B 2012 9 DNA FFPE DNA DNA Input Recommendations DNA 3 DNA Input Recommendations DNA
44. VII 15027983 C JPN Overview dr reese oi SE hi i NE 2 DNA Input Recommendations DNA 3 Additional Resources llana aaao aoaaa LLDD LLALL DDL L LALL DLLD LLL L aLaaa 4 K ee Ge Besse Wees ag O NGGeiearaareeastesrel 7 a TruSeq Custom Amplicon Library Preparation Guide TruSeq Overview Introduction TruSeq TSCA 1 1 536 600 kb TSCA 1 1 536 M
45. 10 2 O Co On 2 Microseal B CLP CLP 1 800 rpm 2 10 CLP 100 Fs Ba KS 2 MK 11 CLP 80 JLT a 80 200 ul
46. MSDSs Ilumina Web www ilumina com msds Product Documentation Humina Web PDF Illuminawww illumina com support Documentation amp Literature TruSeq Custom Amplicon Library Preparation Guide TruSeq p 7 ES ISRAK iAAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACC 3ATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGTTGATCCACTGATTCAACGTACCGTATCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTACCGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGT ENEE EE EE NL e EE Ee iAAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTACC 3ATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACG TGAGACTAAATATTAACGTTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTA ATCAATTGAGACT
47. STAZ SE Gi Ilumina gek 12 Illumina TN KEN De CDR TT ll EE Gv E E Gii E g THE EEROR NEII Gii EE K Illumina E Ki Se E e GE re R 90 est GE o dx SE S So Tlumin Z Illumina
48. Product Files for TruSeq Custom Amplicon TruSeq Custom Amplicon Library Preparation Guide TruSeq6 4 St H L lr L mPOSNIL A9IAJ9AO SS990Jd UO1PJed JIdq AIBIqTT UOoIIdmV WOISND bagna L Supporting Information Prepare Libraries and Sample Sheet MiSeq Ilumina Experiment Manager ID 96
49. 1H 1 8 IJ L VNG SUorJepueunuroo9S Mdu VN y lt AERO E X r Overview Additional Resources TruSeq Illumina support illumina com sequencing kits ilnmn TruSeqW Support Training SN Experienced User Card Lab Tracking Form part 15027985 Illumina Experiment Manager IEM TruSeq TruSeqW Training
50. 20 3 ET IAP TDP1 PMM2 PMM2 TDP1 Procedure 1 45 FPU 2 FPU 2 400 xg 2 3 FPU 50 mM NaOH 25 yl amp NaOH 5 6 a 4 FEPU 5
51. D Protocol Introduction TruSeq 4 Additional Resources Ilumina TruSeq IEM Illumina 4 Additional Resources Ilumina O IEM
52. 0 8 ml MIDI Fisher Scientifc AB 0859 Fisher Scientifc AB 0765 A63881 A63880 Beckman Coulter 538619 Bee C ecend Z REE 200 Ts 5001 Microseal B BIO RAD MSB 1001 PVC Labcor 730 001 250 425 bp Teas 2 150 175 250 bp 4 DNA 1000 Kit Bioanalyzer Agilent 5067 DNA 1504 300 ET TruSeq Custom Amplicon Library Preparation Guide TruSeq Dl Hs tt soqeuwnsuop Doitddnmg Jos var H KS OH E Supporting Information Equipment Pre PCR PCR 37C 96 TY
53. P5 P7 i5 1i7 PCR Estimated Time 85 105 PCR b 30 Consumables PMM2 PCR Master Mix 2 1 15 25C Ilumina 15 i5 A5XX 15 25C Illumina iT A7XX 1 15 25C Illumina TDP1 1 15 25C Illumina TruSeq DNA Polymerase 1 50 mM NaOH 1 10N 96 3 5 NaOH ml Preparation 1 Experienced User Card Custom Amplicon Oligo Tube CAT Lab Tracking Form 8 15027983 KETC JPN PCR
54. Consumables LNA1 Library Normalization 1 15 25C Illumina Additives 1 LNB1 Library Normalization 1 gt Illumina Beads 1 LNW1 Library Normalization 2 2 8C Illumina Wash 1 LNS2 Library Normalization 1 Illumina Storage buffer 2 0 1N NaOH 1 96 3 ml 96 PCR 1 15 ml 1 Microseal B H HE y y E http www illumina com msds MSDS EIN LO TruSeq Custom Amplicon Library Preparation Guide TruSeq 2 0 xb
55. ER ek ee II NAEL Ee E SEN RAL At 12 30 E Illumina Illumina Illumina c e y Ilumina 4 d G Illumina i E ER e Ilumina 1 3 E SC Illumina OH f lumina
56. 20 1 000 xg 1 SHELL TCA 65 HYP FPU FPU 20 FEPU 2 400 xg 2 FPU 15027983 C JPN a _ Stringent Water 1 SW1 45 yl b FPU FPU 2 400 xg 2 JE 2 Stringent Water 1 SW1
57. FPU MIDI H 8 Universal Buffer 1 UB1 45 ul 9 FPU FPU 2 400 xg 2 2 Universal Buffer 1 UB1 10 TruSeq Custom Amplicon Library Preparation Guide TruSeq D Je SO8IIO punoqur JO Teaouay a FHEAE Protocol Extension Ligation of Bound Oligos
58. f K 6 ACD1 ACP1 ID TSCA_Control DNA FFPE TruSeq Amplicon Workflow Specifc Settings Use Somatic Variant Caller CES lumina CAT iT PCR A701 ATCACGAC A702 ACAGTGGT A703 CAGATCCA A704 ACAAACGG A705 ACCCAGCA A706 AACCCCTC A707 cccAAccT A708 CACCACAC A709 GAAACCCA 15027983 C JPN iT PCR AGGGTCAA i5 PCR TGAACCTT TGTTCTCT CTAATCGA TAAGTTCC TruSeq Custom Amplicon Library Preparation Guide TruSeq DAA POSIN UOrTee
59. 0 c o d ei Ilumina Ilumina g Illumina
60. Procedure 1 SGP 2 20 SGP 1 000 xg 1 3 PAL Pooled Amplicon Library 3 4 15027983 CJPN 4 TruSeq Illumina TruSeq CAT e E MiSeq 1 MiSeqv2 v2 MiSeq v3 16 150x 96 500x 96 48 150x 96 500x 96 96 96 150x 96 96 500x 96 96 384 150 96 96 500x 48 80 768 150x 72 96 500x 24 40 1536 150x 36 60 500x 1
61. GoTaq Hot S Start Polymerase 5 338 671 5 587 287 Promega Corporation Promega ji 15027983 C JPN 1 Application Specific IP Ilumina Application Specific IP Illumina Core IP Application Specific IP Core IP Illumina Illumina
62. ACD1 plicon Control DNA 1 3 ACP1 Amplicon Control Oligo Pool 1 OHS2 Oligo Hybridization 15 25C for Sequencing Reagent 2 ELM4 PMM2 TDP1 SW1 e Wash 1 UBI HYP FPU IAP a KEE y geren http www illumina com msds MSDS 4 g 15027983 CJPN 2 2 LNB1 2 8 C LNB1 gt 3 n HT1 Hybridization Buffer
63. Introduction KE EEN tte EES 40 How Does the TruSeq Custom Amplicon Library Preparation Assay Work TruSeq TUTE te OD EEO 41 TruSeq Custom Amplicon Library Preparation Process Overview TruSeq EE d ge E EE 43 CROSS 46 TruSeq Custom Amplicon Library Preparation Kit Contents TruSeq E 48 User Supplied Consumables 51 G DmeT SSSSESRR RM Q EK TOT 550 52 MiSeq Sample Sheet Preparation MiSeq 54 Illumina Amplicon Viewer 56 WW ess a go Y 3 K es ee KN car EE KS z Ze SE Ss SS ad Sen ee uf Be We E At RE a E ae 44receecArearaeAsre67C470 3 a d veg FA e GA ii Ai ro d SEI ep S f Xero f C TruSeq Custom Amplicon Library Preparation Guide TruSeq 0 Supporting Information Introduction
64. 12 DNA Stringent Water 1 SW1 2 Universal Buffer 1 UB1 3 Stringent Water 1 SW1 y Y ET http www illumina com msds MSDS UI r AI EEN B
65. DesignStudio 2 50 mM NaOH 1 3 Illumina Experiment Manager TruSeq Experienced User Card 4 15 25 PCR Master Mix 2 PMM2 G5 i7 PCR Master Mix 2 PMM2 20 5 1 7 mL 4 Additional Res
66. Estimated Time 1 35 15 EI Consumables CAT Custom Amplicon Oligo 1 15 25C Illumina Tube OHS2 Oligo Hybridization 1 FD z n Illumina for Sequencing 2 ACD1 1 15 25C Illumina ACP1 T li Illumina DNA 5 3 DNA Input Recommendations DNA 96 PCR lzi 2 9 15027983 CJPN Preparation 1 3 DNA Input Recommendations DNA 15 25 Custom Amplicon Oligo Tube CAT Oligo Hybridization for Sequencing 2 OHS2 Amplicon Control DNA 1 ACD1 Amplicon Control Oligo Pool 1 ACP1 DNA DNA
67. A260 A280 1 8 2 0 E8520 DNA Assessing DNA Quality DNA FFPE DNA TruSeq FFPE DNA OC Ilumina FFPE QC WG 321 1001 A Cq lt 2 0 UV H dsDNA 260 OD RNA D Illumina FFPE QC 260 nm 280 nm G 2 0 DNA TruSeq Custom Amplicon Library Preparation Guide TruSeq
68. S 95C 3 X 95 30 66 30 72C T 60 72C 5 10C SAFE STOPPING POINT d 24 PCR Clean Up PCR oO LC EE Et onbe 1 2 8C 2 2 8 Microseal AT Microseal B TruSeq Custom Amplicon Library Preparation Guide TruSeq 2 E EI VM UOeoJIdmV yod OI Protocol PCR Clean Up PCR PCR AMPure XP Beads Estimated Time gt 50 b 20 Consumables EBT Elution Buffer Tris 1 Illumina AMPure XP Beads EE 80 96 40 EtOH ml 96 M
69. 5 EPU PMM2 TDPI 22 ul IAP 6 FPU a P20 20 il b HH IAP JLT FPU 1 NaOH 5 6 FPU NaOH 20 u IAP E 5 6 FPU DNA PMM2 TDPI FPU c _ FPU
70. Application Specific IP Illumina Application Specific IP 3 Ee 4 a b Illumina Illumina
71. Oligo Hybridization for Sequencing 2 OHS2 96 95 37 Experienced User Card Ilumina Experiment Manager TruSeq Amplicon TruSeq DNA CAT ACD1 ACP1 TSCA_ControD Illumina 16 1
72. DNA FFPBE DNA 10 u 25 ng l DNA 15 d DNA DNA FFPB DNA DNA 250 ng 25 ng ul 250 ng gt 16 7 ng ul 4 Amplicon Control DNA 1 ACD1 Amplicon Control Oligo Pool 1 ACP1 5 yl 5 DNA Custom Amplicon Oligo Tube CAT 5 T 70 8 PCR Custom Amplicon Oligo Tube CAT 12 Illumina 6
73. SGP StoraGe Plate 96 PCR SGP Library Normalization Storage Buffer 2 LNS2 30 ul Zi 5 LNP 15027983 C JPN EA RS 1 800 pm 5 17 LNP 2 18 30 LNP SGP
74. 3 FPU L 4 4 Additional Resources 2 TruSeq Custom Amplicon Library Preparation Guide TruSeq 3 Je so31O punoquq JO Teaouay caL FH Protocol B C D MIDI FPU Filter Plate Unit FPU FPU a Stringent Water 1 SW1 45 ul b FPU c 20 FPU 2 400 xg 10
75. 18 PCR Amplification PCR 24 PCR Clean Up PCR 15027983 2011 10 5 TruSeq Custom Amplicon Library Preparation Guide TruSeq vii 201 ATO SIH UOISIASS H EE Table of Contents Revision History vi Table ofContents EE Ee ee viii 1 Overview 1 Introduction aaaaaaaaaaa0aaa00a 00000000000001 BC 2 DNA Input Recommendations DNA 3 Additional Resources ollaaa aaao oaaao hLDL LLLA aa LDLo aLaaa aooo 4 2 Protocol 5 Introduction aaan PRE E E E eee 6 TruSeq Workflow TruSeq LL EE 7 Hybridization of Oligo Pool 8 Removal of Unbound Oligos LL 12 Extension Ligation of Bound Olig
76. Extension Ligation Mix 4 Hybridization Buffer Indexed Amplification Plate Library Normalization Beads 1 LNS2 Library Normalization Storage Buffer 2 OHS2 Oligo Hybridization for Sequencing Reagent 2 4 6 15027983 C JPN PCR Master Mix 2 Stringent Wash 1 Universal Buffer 1 TruSeq Custom Amplicon Library Preparation Guide TruSeq 47 SUUAUOJOV f X ke Sa naf Supporting Information TruSeq Custom Amplicon Library Preparation Kit Contents TruSegq TruSeq EE TruSeq Custom Amplicon Library Preparation Kit v1 5 Catalog FC 130 1001 TruSeq v1 5 FC 130 1001 1
77. 570b 3 ACP1 ACD1 DCH 1500 1000 700 500 400 300 200 100 A 250 bp PCR 350 bp B TruSeq Custom Amplicon Library Preparation Guide TruSeq D G Bh co Ad IN Ad Protocol 4 Bioanalyzer ACP1 ACD1 PCR FU i r 3 TT CT T CT T 15 100 200 3200 500 1500 bp A D 250 bp PCR 350 bp C TruSeq CAT Bioanalyzer Ilumina Amplicon Control DNA 1 ACD1 Amplicon Control Oligo Pool 1 ACP1
78. CLP 10 CLP Elution Buffer Tris EBT 30 Microseal B CLP 1 800 rpm 2 2 2 2 LNP Li
79. DNA DNA DNA ELET 5 DNA DNA PCR DNA Estimated Time 50 5 Consumables ELM4 Extension Ligation Mix 4 1 IR 25C 1 Procedure Illumina 1 FPU ELM4 45 ul
80. LNA1 LNWI LNS2 EBT CLP LNP SGP PAL DAL b 4 TruSeq CAT Custom Amplicon Oligo Tube 15 25 C TruSeq Custom Amplicon Library Preparation Index Kit Catalog FC 130 1003 TruSeq FC 130 1003 gt 1 i5 A501 A508 8 i7 A701 A712 12 gt 2 i5 i7 TruSeq Custom Amplicon Library Preparation Guide TruSeq 49 a DI LGS Mrolc keng jmD9STIT S1U91UO Fy UOIB Bd9JT AIEIqTT UO9IIdOV UmO1Sn
81. MiSeq MiSeq Reporter MiSeq Reporter TruSeq bam vcf Seg System User Guide MiSeq MiSeq Reporter http www illumina com help miseq_reporter default htm Illumina Amplicon Viewer Illumina Amplicon Viewer TruSeq TSCA Amplicon Viewer MiSeq Q DesisnStudio
82. 1 2 P5 A B C PCR D MiSeq TruSeq Custom Amplicon Library Preparation Guide TruSeq 4 DI al LAC LTL IL mbSNIT YOM ASSV uorpTedosg ABJdTT UOOIIdmV wozsno bagna g 94 S9o MOH E EE EEO Le GE HJN 01 uoneuiojul sunsoddng JC JPN R 15027983 42 TruSeq Custom Amplicon Library KE Process Overview TruSeauWw TruSeq Design Your Project Ilumina DesignStudio DesignStudio
83. 1 LNA1 15027983 C JPN 3 Library Normalization Beads 1 LNB1 SERAL D Eb P1000 800 ul Library Normalization Additives 1 LNA1 15 ml 15 20 LNA1 LNB1 96 96 2 1 000 ul P1000 3 Library Normalization Additives 1 LNA1 Library Normalization Beads 1 LNB1 P200
84. VWR Internationa Scigene Hybex Microsample Incubator PCR PCR vw E d Si Y ATH o tt Post PCR PCR Magnetic Stand 96 PCR Invitrogen DynaMag 96 Side Skirted Q Instruments BioShake iQ 1808 0506 BioShake XP 1808 0505
85. boenil Supporting Information 50 Additional Components TruSeq RE Illumina FFPE OC FFPE DNA TruSeq FC 130 1005 dite TruSeq FC 130 1007 15027983 CJPN User Supplied Consumables 3 3 3 2 40 ml 2 10 N NaOH 96 PCR 0 2 ml BIO RAD MSP 9601 96
86. 2 20 Custom Amplicon Oligo Tube CAT ER 5 SGP P200 3 5 6 P20 5 1 SGP 8 PCR Microseal B SGP BEL SS i EH HY TruSeq Custom Amplicon Library Preparation Guide TruSeq D LLAD POSIN OP ELL LO ALL LA Upeo 9 dOeS D9SIIA pue Suooq AJeJdTT Protoco
87. 30 30 7 2 8 LNP 80 ul 2 Ne 9 LNP Library Normalization Wash 1 LNW1 TruSeq Custom Amplicon Library Preparation Guide TruSeq d L t L HOUSEN Aq Protocol 12 13 14 a _
88. AA TA OT A CAA SE A A ANRA AE AA LAL wem JA ATTA G GCA AC AAGCTIAAGATTACTTGAI EE AACCTI EE El CTGTTIAACCTTAAGATTACTTGATCCAC GAAAAGAATG 3AAAAGAATGAT AACAGTAAOACACTTCTGTTAACCTD AAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGAT IIAAGAGCTAC EE ATCAATTGAGACTA CACACTTCTGT CCAT TAAGAGCTACCGTGCAACAGTAACACACTTCTGTTAAC REN A GAA AA GAT GATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAA ee TA GTACCG ACGAACGTATOALTAAGATTACTTGATCCACTGATTCAACGTA CCGTAACGAACGTA CAATTGAGA CTAAA TGTTAACCTT GTTGATCCAGTGATTCAACGT IAAGATTACTTGATCCACTGATTCAACGTACCG TAACGAACG TATCAATTGAGCTTCTGTIAAG TAGCAACGACG AAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGAT d TAAGAGCTACC FOR RESEARCH USE ONLY ILLUMINA PROPRIETARY FC 130 9005DOC 15027983 C JPN 2013 8 Illumina Inc Illumina Illumina
89. AAATAT TAACGTACTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAACGACGA 3ACTAAATAT TAACGTACCAT TAAGAGCTACAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTGCAACGACGAAAAGAATGATAACAGTAACACA ANOACLL E El EE E EE NEE CCAT TAAGAGCTACCGTGCAACTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACCGTGCAACGACGAACTTCTGTTAACCT TAAGATTACTTGAT 3CTACCGTGCAACGAAAATAACCTTAAGATTACTTGATCCACTGATTCAACGTACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAGCTACCGTGCAACGACGAAAAGAATGAT Ate e EE RACO TIAAGAT EE E E NEE EE 3ATAACAGTAACACACTTCTGTTAACCT II A LE OR TCAATTGAGACT ees CGACGAAAAGAATGA Uer CCAT TAAGAGCTACCGTGCAACAGTAACACACTTCTGT TAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACCGTGCAACGACGAAAAGAATGATAA SAACAGTAACACACTTCTGTIAACCTIAAGALIACTIGATCCACTGATTOAACGIACOG1AACGAAOG ATCAATTGAGACTAAATAI TAACGTACOAIAAGAGCTAOGTC11CTGT IAACCTIAAGATIACT1GATCCAOTGAITOAACG GTACCGTAACGAACGTATCATTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTG CCGTGCAACGACGAAAAGAATGATAACAGTAACACACTTCTGT TAACCTTA gt TTGATCCACTGATTCAACGTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGCTTCTGTTIAACCTTAAGATTACTT ICAO TGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAGCAACGACG AAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCAC ele GTAACGAACGTATCAATTGAGACTAAATATTAACGTACCAT
90. ACTGATTCAACGTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACG TACCGTAACGAAOGTALCAATTGAGA CTAGCAACGACG e EE E EE eege Sen Ee e CCGTGCAACGACGAAAAGAATGATAACAGTAACACACTTCTGT TAACCTT CTTGATCCACTGATTCAACGTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTA TCAATTGAGACTAGCAACGACG GE E ee E E E GATAACAGTAACACACTTCTGTTAACCTTAAGAT TL CTAAATATTAACGTACCATTAAGAGCTACCGTCTTCTGTTAACCTTAAGATTACTTGATCCACT GE CCGT CACTGATTCAACGTACCAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTCTTCTGT TAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGA SAAAAGAATGATAACAGTAACACAC TTCTGTTAACCTIAAGAACTTGATOCACTGALTCAACGTAGOG TAAAGATIACTTGATCCACTGATTCAACG TACCGTAACGAACGTATCAATTGAGAGTAAATATTAACGTACCAT 1AAGAGOTACX UA OR RO RS Ee CAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTCTTCTGTTAACCT TAAGATTACTTGATCCACTGATTCAAC TTGAGACTAAATATTAACGTTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTTCTGTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTA 1AICAATTGAGAOTAAAIALIAAOG1ACTIAACOTIAAGALIACTTGATCOAGTGAITOAAOGTACOG1AACGAACGTOTTCTGHAACOIAAGATIACTTGATCOAOTGATICAAOGIACCGTAACGAACGTIATOAAT1 GAGAG TAC GAGGA GACTAAATATTAACGTACCAT TAAGAGCTACAACCTTAAGATTACTTGATCCACTGATTCAACGT EE TCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACCGTGCAACGACGAAAAGAATGATAACAGTAACAC GATAACAGTAACACACTTCTGTTAACCT TAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGAC AAA TAA AC TT TAA
91. ACTTCTGTTAACCT TTACTTGATOCACTGAT TOAACGT IACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGT ACCATTIAAGAGCTACCGTGCAA AOGA CSAAAAGAA TGA AAACAG1AACACACTTCTGT1 CCATTAAGAGCTACCGTEGCAACAGTAACAGACTTCTGTTAACCT TAAGATTACTTGATCCACTGATTCAACGTACCG TAACGAACGTATCAATTGAGACTAAATATIAACGTAGCATTAAGAGCTACCGTGOAACGACGAAAAGAATGATAA 3ATAACAGTAACACACTTCTGTL AAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACCGTCTTCTGTTL IAAGAT TI KEE e gaTACCGT AACGAACGTATCATTAAGATTACTTGATOCACTGAITCAACGTA CCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTGCAACGA OGAAAAGAA GENY TAACAGTAACACACTTCTGTTAACCTT PTTGATCCAGTGATTOAAOGTTAAGALAOTTGATGOAOTGATTOAAOGTAGOGTAAOGAAOGTATOARLTGAGCTTCTGTTAAOOTTAAGATTACTTGATCCACTGATTCAAOGTAOCGTAACGAAOGTATOAALTGAGACTAGCAAOGAOG iAAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTACC Illumina San Diego California 92122 U S A 1 800 809 ILMN 4566 1 858 202 4566 techsupport ilumina corm www ilurmina corm
92. CTGATTCAACGTACCAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACCGTCTTCTGTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAM AAAAGAATGAIAACAGTAAOACACTICTGTIAACCTTAAGATIACTTGATGCACTGATTCAACGTACCGTAAAGAIACTTGATCCACTGAITCAACGIACOGTAAGGAAOGTATOAATTGAGACTAAAIATTAACGTACGATIAAGAGCTACC EE E GTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAA Mee ee EE ACTTGATCCACTGATTCAACG TGAGACTAAATATTAACGTTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATA UE GCTTCTGTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAM ATCAATTGAGA CTAAATATL AACGTACTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTCTTCTGTTAACCTTAAGATT AGTTGATCCAGTGATTCAACGL ACCGTAACGAACGTATCAATTGAGACTAACGACGA AAACAGTAACAOACLTGTGT IAACOTIAAGATTIACTIGATCCAGTGATTOAACGTAOOGTAAOGAACG1AIOAAITGAGAGTAAATATTIAAOGTACOAHTAAGAGCTACOGTCTTCTGTTAACOLUAAGALIACTTGAICCACTGAI 1OAACG EA 3ATAACAGTAACACACTTCTGT TAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTA TTCTG CCTTAAGATTACTTGATCCACTGA CCATTAAGAGCTACCGTGCAACTTAACCTTAAGATTACTTGATCCACTGAT TCAACGTACCGTAACGAACGTATCAAT TGAGACTAAATAT TAACGTACCAT TAAGAGCTACCGTGCAACGACGAACTTCTGTTAACCTTAAGATTACTTGAT CTACCGTGOAAOGAAAATAACO TIAAGATIACTTGATCCACTGATTCAACG TACTTCTGTTAACCTTAAGATTACTTGATCCACTGATOAACGTACCGTAACGAACGTATCAATTGAGACTAAGCTACCGTGCAACG ACGAAAAGAATGAT iAAAAGAATGA ee AL EE GAT TACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGAT MR EN UE AACGAACGTA TTGAGACTAAA CGTACCATTAAGAGCTACC 3ATAACAGTAACAC
93. Custom Amplicon Library Preparation Guide TruSeq dc HOUSEN ARIT Protocol Library Pooling and MiSeq Sample Loading MiSeq MiSeq TN Hybridization Buffer HT1 Estimated Time 10 b 10 Consumables HT1 Hybridization Buffer ina PCR Dq 2 5L Preparation 1 1 5ml 96 2 15 25C MiSeq 3 3 1
94. IDI 2 BEE e Microseal D Preparation 1 AMPure XP Beads 2 80 der 80 80 S Procedure 1 20C IAP 1 000 xg 1 4 15027983 CJPN 3 PCR 4 5 ul Bioanalyzer 1 ui PCR PCR 150 bp 280b 175 bp 310b 250 bp 350b 425 bp
95. Ilumina Experiment Manager Windows Illumina Experiment Manager Ilumina SE www illumina com TruSeq Downloads MyIllumina Sequence Libraries on MiSeq MiSeq TruSeq MiSeq MiSeq MiSeq System User Guide MiSeq Analyze Data with Illumina Supported Software Illumina MiSeq Reporter MiSeq Reporter
96. La O LL L HOUSEN Aq Protocol y B ZC ET Preparation 4 Additional Resources 0 1N NaOH 1 25C Library Normalizatio 250C 15 207 Library Normalizati VET ormalization Beads 1 LNB1 Library Normalization 2 8 Wash 1 LNW1 TERJED D Library 207 JELET 25
97. M4 HT1 FPU PAL amp DAL CV PCR 30 85 105 B aen TDP1 ZE es Sep SE JT i7 NaOH IAP CL 4 TruSeq Custom Amplicon Library Preparation Guide TruSeq mPOSNIL D AC CO Ath ef EC wu DOT LU MOPAIOM gg D i Protocol Hybridization of Oligo Pool DNA y Ell RESE AET So TOD http www illumina com msds MSDS
98. TAAGAGCTACC 3ATAACAGTAACACA LIU AG CCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAAC ALTAAGAGC1ACCGTOTTCTGT IAA CCTTAAGATTACTTGA e EE Se TCATTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACCGTGCAACGACGAAAAGAATGATAACAGTAACACACTTCTGTTAACCT iAAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAAT ATMAOGTACCATTAAGAGCTACC CCATTAAGAGCTACCGTGCAACAGTAACACACTTCTGT TAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTGCAACGACGAAAAGAATGATAA 3ATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCAT TAAGAGCTACCGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACG ZTTGATCCACTGATTCAACGT TAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGCTTCTGT TAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAGCAACGACG GE EE E e ee STACCGTAACGAACGTATCATTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGAN ee GE CGACGAAAAGAATGATAACAGTAACACACTTCTGT TAACCT II gt TTGATCCACTGATTCAACGTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGCTTCTGTTIAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATC AATTGAGACTAGCAAGGACG AAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACC 3ATAACAGTAACACACTTCTGTTAACCT TAAGAT TACTT Ee AAATATTAACGTACCATTAAGAGCTACCGTCTTCTGTTAACCTTAAGAT TA Lt yA
99. TGTTAACCTL AAGATAOTTGATCCACTGATTCAAOGTACCGTAACGAA CGTATCAAI NTGAGACIAAGCTACOGTGCAACGAN CGAAAAGAATGA E EE NOIMAOSAACOTUCAAU GALKVAIALIAACGIACSALIAAGAGCNAKA GATAACAGTAACACACTTCTGTTAACCTTAAG E ACCGTAACGAACGTATCAATTGAGAC ee CGACGAAAAGAATGA EH ACCATTAAGAGCTACCGTGCAACAGTAACACACTTCTGTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTACCGTGCAACGACGAAAAGAATGAT GAIAACAGTAAOACACTTCGTIAACCTIAAGA AGITGATOOACTGAITCAAOGTACCG1AACGAAOGTAICAATTGAGACTAAAIA IAAOGTAOCATIAAGAGCTACCGTCTTCTG IDAAOCTIAAGATTACTTOAIOOAOTGAT CA Ee ee CCGTGCAACGACGAAAAGAATGATAACAGTAACACACTTCTGTTAACCTT CTTGATCCACTGATTCAACGTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTA TCAATTGAGACTAGCAACGACG 3AAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTAC GATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGA RA CGTACCATTAAGAGCTACCGTCTTCTGTTAA ed EE EE NEE GTACCGTAACGAACGTATCA AE ge ACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCAT TAAGAGCTACCGTGCAACGACGAAAAGAATGATAACAGTAACACACTTCTGTTAACCTT 3AAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAAAGAT TACTTGATCCACTGATTCAACGT ACCGIAAOGAACGTATCAATTGAGACTAAAIAT IAACGT ACCATTAAGAGCTACC ee E E EE E Ee GATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGT Eed ACCATTAAGAGCTACCGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAAC CTTGATCC
100. brary Normalization Plate MIDI ET P20 CLP 20 ul VEL LNP 2 ER Microseal B LNP 1 000 xg 1 SELLET 15027983 C JPN Library Normalization Estimated Time 1 20 30
101. de 199HS 9Idue baSIA Supporting Information Illumina Amplicon Viewer D6 MiSeq MiSeq Reporter Illumina Amplicon Viewer Amplicon Viewer TruSeqW SCA Amplicon Viewer MiSeq Q Amplicon Viewer MiSeq Reporter Amplicon Viewer MyIllumina http www ilumina com help amplicon_viewer default htm Illumina Ampl
102. iSeq 1 96 TSCA MiSeq SR Excellent Multiplexing Capability 1 1 536 1 MiSeq 96 Revolutionary Assay with Fast and Simple Workflow 3 1 96 1 536 Customized Design Illumina DesignStudio Automated Data Analysis
103. icon Viewer TruSeq Training 15027983 C JPN Ilumina 5 Ilumina Illumina Web 6 Illumina 1 800 809 4566 800 874909 10800 296575 0800 0223859 0800 81102 800 16836 80882346 900 812168 0800 918363 020790181 10800 911850 0800 563118 0800 180 8994 70 0800 917 0041 1800 812949 44 1799 534000 MSDSs D
104. illumina TruSeoW 3AAAAGAATGATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAACGTACCATTAAGAGCTACC GATAACAGTAACACACTTCTGTTAACCTTAAGATTACTTGTTGATCCACTGATTCAACGTACCGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGT NCAA Ee ACCGTAACGAACGTA El Kee EH ACCATTAAGAGCTACCGTCTTCTGTTL Kee GAT TACTTGATCCACT E Ek ACCGTAACGA AACAGTAACACACTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTAN MTACTTGATCCACTGATTCAACGTACCGTAACGAAN TCAATTGAGACTAAATATTAACGTACCAT TAAGAGCTACC AAACAG TAACACACTTCTG TIAACCTIRAGATTACTITGATCCACTGATTCAACGTACCG AACGAACGTATGAATTGAGACTAAATATIAACGTACCATTAAGAGCTACCOGTCTTOTGT TAACCT TAAGALIACTTGATCCACTGATTCAACC TTGAGACTAAATATTAACGTTGTTAACC de VE GATCCACTGATT EACCR ACGAACA er GACTAAATATTAACGTACCATTAAGAGCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTA TATCAATTGAGA ACTAAATALTAACGT IACTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTCTTCTGTTAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAACGACG GACTAAATAT TAACGTACCATTAAGAGCTACAACCTTAAGATTACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATATTAACGTACCATTAAGAGCTACCGTGCAACGACGAAAAGAATGATAACAGTAACAC Ee E e E e TORN ENAC E EE e E el EAA ACCATTAAGAGCTACCGTGCAACTTAACCTTAAGAT TACTTGATCCACTGATTCAACGTACCGTAACGAACGTATCAATTGAGACTAAATAT TAA ee CGACGAACTT AACCTTAAGATTACTTGAI GCTACCGTGCAACGAAAATAACCTTAAGATTACTTGATCCACTGATTCAACGTACTTC
105. l 4 Additional Resources p SGP 15 25 8 PCR PAL Pooled Amplicon Library PAL DAL Diluted Amplicon Library HT1 MiSeq 3 PAL DAL PAL 3 5 PAL HT1 20 25C
106. os 16 DCH Amplification DCH 18 PCR Clean Up PCR EE EE 24 Library Normalization eegesee e 29 Library Pooling and MiSeq Sample Loading MiSeq ee EE 34 A Supporting Information 39 Introduction osceann eeaeee a AAAA AAAI EA Enan 40 How Does the TruSeq Custom Amplicon Library Preparation Assay Work TruSeq EE 41 TruSeq Custom Amplicon Library Preparation Process Overview TruSeq 43 Acronyms CRT aa EE 46 TruSeq Custom Amplicon Library Preparation Kit Contents TruSeq oaoieeai didoeroek Aei eai aibre aed aulai eai 48 User Supplied Consumables 51 ENT DE EE 52 MiSeq Sample Sheet Preparation MiSeq 54 Illumina Amplicon Viewer laaa aaao oaaao aaao aooaa LaLa LLDD DLL Da aa Laaa 56 TL I ER 57 TruSeq Custom Amplicon Library Preparation Guide TruSeq
107. ources 6 TruSeq a 5 H b i7 1 12 TruSeq Custom Amplicon Library Preparation Guide TruSeq 0 d r Od UOeoIIdmV yod H D Protocol 20 9 A ij5 B iT C IAP IAP Indexed Amplification Plate 96 PCR IAP ij5 4 ul
108. uencing Reagent 2 OHS2 Custom Amplicon Oligo Tube CAT 9 20C 1 000 xg 1 10 95 HYP 1 11 40 80 K 95 40 C EE 80 PCR TruSeq Custom Amplicon Library Preparation Guide TruSeq GG Lena keete LE E Od OSHO JO UOTNRZIPIIqGAH Protocol Removal of Unbound Oligos
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