14 GeneChip ® <strong>Mapping</strong> <strong>500K</strong> Assay Manual 42. Klein, R.J., Zeiss, C., Chew, E.Y., Tsai, J.Y., Sackler, R.S., Haynes, C., Henning, A.K., Sangiovanni, J.P., Mane, S.M., Mayne, S.T., Bracken, M.B., Ferris, F.L., Ott, J., Barnstable, C., Hoh, J. Complement factor H polymorphism in age-related macular degeneration. Science 308:385-9 (2005). 43. Serono Identifies 80 Genes Involved in Multiple Sclerosis Using 100,000 <strong>SNP</strong>s. In: Affymetrix Microarray Bulletin; 2005; Issue 1: 1- 4; www.microarraybulletin.com. 44. Huang, J., Wei, W., Zhang, J., Liu, G., Bignell, G.R., Stratton, M.R., Futreal, P.A., Wooster, R., Jones, K. W., Shapero, M.H. Whole genome DNA copy number changes identified by high density oligonucleotide arrays. Hum Genomics 1:287-99 (2004). 45. Nannya, Y., Sanada, M., Nakazaki, K., Hosoya, N., Wang, L., Hangaishi, A., Kurokawa, M., Chiba, S., Bailey, D. K., Kennedy, G.C., Ogawa, S. A robust algorithm for copy number detection using high-density oligonucleotide single nucleotide polymorphism genotyping arrays. Cancer Res 65:6071-9 (2005). 46. Wong, K.K., Tsang, Y.T., Shen, J., Cheng, R.S., Chang, Y.M., Man, T.K., Lau, C.C. Allelic imbalance analysis by high-density single-nucleotide polymorphic allele (<strong>SNP</strong>) array with whole genome amplified DNA. Nucleic Acids Res 32:e69 (2004). 47. Zhao, X., Li, C., Paez, J.G., Chin, K., Janne, P.A., Chen, T.H., Girard, L., Minna, J., Christiani, D., Leo, C., Gray, J.W., Sellers, W.R., Meyerson, M. An integrated view of copy number and allelic alterations in the cancer genome using single nucleotide polymorphism arrays. Cancer Res 64:3060-71 (2004). 48. Zhou, X., Mok, S.C., Chen, Z., Li, Y., Wong, D.T. Concurrent analysis of loss of heterozygosity (LOH) and copy number abnormality (CNA) for oral premalignancy progression using the Affymetrix 10K <strong>SNP</strong> mapping array. Hum Genet 115:327-30 (2004). 49. Rauch, A., Ruschendorf, F., Huang, J., Trautmann, U., Becker, C., Thiel, C., Jones, K.W., Reis, A., Nurnberg, P. Molecular karyotyping using an <strong>SNP</strong> array for genomewide genotyping. J Med Genet 41:916-22 (2004).
chapter 1 | Overview 15 50. Bignell, G.R., Huang, J., Greshock, J., Watt, S., Butler, A., West, S., Grigorova, M., Jones, K.W., Wei, W., Stratton, M.R., Futreal, P.A., Weber, B., Shapero, M.H., Wooster, R. High-resolution analysis of DNA copy number using oligonucleotide microarrays. Genome Res 14:287-95 (2004). 51. Janne, P.A., Li, C., Zhao, X., Girard, L., Chen, T.H., Minna, J., Christiani, D.C., Johnson, B.E., Meyerson, M. High-resolution single-nucleotide polymorphism array and clustering analysis of loss of heterozygosity in human lung cancer cell lines. Oncogene 23:2716-26 (2004). 52. Herr, A., Grutzmann, R., Matthaei, A., Artelt, J., Schrock, E., Rump, A., Pilarsky, C. High-resolution analysis of chromosomal imbalances using the Affymetrix 10K <strong>SNP</strong> genotyping chip. Genomics 85:392-400 (2005). 53. Lieberfarb, M.E., Lin, M., Lechpammer, M., Li, C., Tanenbaum, D.M., Febbo, P.G., Wright, R.L., Shim, J., Kantoff, P.W., Loda, M., Meyerson, M., Sellers, W.R. Genome-wide loss of heterozygosity analysis from laser capture microdissected prostate cancer using single nucleotide polymorphic allele (<strong>SNP</strong>) arrays and a novel bioinformatics platform dChip<strong>SNP</strong>. Cancer Res 63:4781-5 (2003). 54. Ishikawa, S., Komura, D., Tsuji, S., Nishimura, K., Yamamoto, S., Panda, B., Huang, J., Fukayama, M., Jones, K.W., Aburatani, H. Allelic dosage analysis with genotyping microarrays. Biochem Biophys Res Commun 333:1309-1314 (2005). 55. Zhao, X., Weir, B.A., LaFramboise, T., Lin, M., Beroukhim, R., Garraway, L., Beheshti, J., Lee, J.C., Naoki, K., Richards, W.G., Sugarbaker, D., Chen, F., Rubin, M.A., Janne, P.A., Girard, L., Minna, J., Christiani, D., Li, C., Sellers, W.R., Meyerson, M. Homozygous deletions and chromosome amplifications in human lung carcinomas revealed by single nucleotide polymorphism array analysis. Cancer Res 65:5561-70 (2005). 56. Garraway, L.A., Widlund, H.R., Rubin, M.A., Getz, G., Berger, A.J., Ramaswamy, S., Beroukhim, R., Milner, D.A., Granter, S.R., Du, J., Lee, C., Wagner, S.N., Li, C., Golub, T.R., Rimm, D.L.,
- Page 1 and 2: GeneChip ® Mapping 500K Assay Manu
- Page 3 and 4: Contents CHAPTER 1 Overview 1 ABOUT
- Page 5 and 6: contents v Equipment and Consumable
- Page 7 and 8: contents vii Reagents Required 118
- Page 9 and 10: contents ix CHAPTER 8 Troubleshooti
- Page 11 and 12: contents xi STEP 4: PCR 257 Reagent
- Page 13 and 14: Chapter 1 Overview
- Page 15 and 16: About This Manual This manual is a
- Page 17 and 18: chapter 1 | Overview 5 LD patterns
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- Page 21 and 22: chapter 1 | Overview 9 Gilman, B.,
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- Page 31 and 32: Chapter 2 Laboratory Setup
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- Page 35 and 36: Main Lab Pre-PCR Clean Room chapter
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About Controls chapter 4 | 96-Well
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B P1 P2 P3 Figure 4.2 Transferring
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Table 4.16 PCR Master Mix ADD PCR M
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chapter 4 | 96-Well Plate Protocol
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WHAT YOU CAN DO NEXT chapter 4 | 96
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EQUIPMENT AND CONSUMABLES REQUIRED
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POOL THE PCR PRODUCTS chapter 4 | 9
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WHAT YOU CAN DO NEXT chapter 4 | 96
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REAGENTS REQUIRED chapter 4 | 96-We
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chapter 4 | 96-Well Plate Protocol
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Figure 4.5 Loading Optical Plates w
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Process Control Metrics Evaluate th
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Table 4.24 PROBLEM: Average Sample
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WHAT YOU CAN DO NEXT chapter 4 | 96
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EQUIPMENT AND CONSUMABLES REQUIRED
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IMPORTANT INFORMATION ABOUT THIS ST
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Figure 4.6 Work Area Set Up for Sta
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chapter 4 | 96-Well Plate Protocol
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WHAT YOU CAN DO NEXT chapter 4 | 96
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Stage 8: Labeling ABOUT THIS STAGE
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REAGENTS REQUIRED chapter 4 | 96-We
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chapter 4 | 96-Well Plate Protocol
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WHAT YOU CAN DO NEXT chapter 4 | 96
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LOCATION AND DURATION • Main Lab
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Hybridizing Samples Using Heat Bloc
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chapter 4 | 96-Well Plate Protocol
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chapter 4 | 96-Well Plate Protocol
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Table 4.37 Hybridization Master Mix
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Load the Samples onto Arrays chapte
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Figure 4.8 Applying Tough-Spots ®
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chapter 4 | 96-Well Plate Protocol
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chapter 4 | 96-Well Plate Protocol
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Chapter 5 Washing, Staining, and Sc
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Introduction This chapter contains
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Reagent Preparation Wash A: Non-Str
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Experiment and Fluidics Station Set
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chapter 5 | Washing, Staining, and
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chapter 5 | Washing, Staining, and
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chapter 5 | Washing, Staining, and
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chapter 5 | Washing, Staining, and
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HANDLING THE GENECHIP ® PROBE ARRA
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chapter 5 | Washing, Staining, and
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Chapter 6 Fluidics Station Care and
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Introduction This chapter provides
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chapter 6 | Fluidics Station Care a
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chapter 6 | Fluidics Station Care a
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chapter 6 | Fluidics Station Care a
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chapter 6 | Fluidics Station Care a
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chapter 6 | Fluidics Station Care a
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White Clamp Figure 6.7 Releasing pe
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Figure 6.8 Troubleshooting decision
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PROBLEMS AND SOLUTIONS Table 6.3 Co
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Table 6.3 (Continued) Common error
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Table 6.4 (Continued) Common Error
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Table 6.5 (Continued) Other Problem
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Chapter 7 Analysis Workflow
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Introduction Software Requirements
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Figure 7.2 GTYPE main window with t
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Figure 7.3 Dynamic Model Mapping Al
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Table 7.1 Dynamic Model Mapping Alg
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FILE SETS chapter 7 | Analysis Work
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chapter 7 | Analysis Workflow 191 3
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NETAFFX SNP ANNOTATION chapter 7 |
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CALL RATE chapter 7 | Analysis Work
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chapter 7 | Analysis Workflow 197 G
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chapter 7 | Analysis Workflow 199 F
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chapter 7 | Analysis Workflow 201 S
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B2 OLIGO PERFORMANCE chapter 7 | An
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chapter 7 | Analysis Workflow 205 3
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chapter 7 | Analysis Workflow 207 W
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Chapter 8 Troubleshooting
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Assay Recommendations 211 Genotypin
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chapter 8 | Troubleshooting 213 10.
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Problem Likely Cause Solution Faint
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Problem Likely Cause Solution Posit
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Table 8.2 PROBLEM: Average Sample O
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When to Contact Technical Support c
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Appendix A Reagents, Equipment, and
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Reagents, Equipment, and Consumable
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appendix A | Reagents, Equipment, a
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appendix A | Reagents, Equipment, a
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appendix A | Reagents, Equipment, a
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Consumables Required GENECHIP ® AR
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Supplier Contact List Table A.10 Su
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Appendix B Thermal Cycler Programs
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ABOUT THIS APPENDIX 500K DIGEST 500
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500K FRAGMENT 500K LABEL 500K HYB a
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Appendix C Low Throughput Protocol
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Introduction The Affymetrix GeneChi
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BEFORE YOU BEGIN appendix C | Low T
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STEP 2: Restriction Enzyme Digestio
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DIGESTION PROCEDURE PRE-PCR CLEAN A
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STEP 3: Ligation REAGENTS AND EQUIP
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PCR STAGING AREA appendix C | Low T
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STEP 4: PCR REAGENTS AND EQUIPMENT
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PCR PROCEDURE PRE-PCR CLEAN ROOM ap
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MAIN LAB appendix C | Low Throughpu
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STEP 5: PCR Purification and Elutio
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appendix C | Low Throughput Protoco
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STEP 7: Fragmentation REAGENTS AND
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FRAGMENTATION PROCEDURE 1. Pre-heat
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appendix C | Low Throughput Protoco
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appendix C | Low Throughput Protoco
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LABELING PROCEDURE MAIN LAB appendi
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STEP 9: Target Hybridization REAGEN
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HYBRIDIZATION PROCEDURE appendix C
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appendix C | Low Throughput Protoco
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Appendix D Reagents, Instruments, a
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Appendix D
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appendix D | Reagents, Instruments,
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* Adaptor, Nsp 5' ATTATGAGCACGACAGA
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Table D.4 Reagents Supplied by Affy
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appendix D | Reagents, Instruments,
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appendix D | Reagents, Instruments,
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Fragmentation and Labeling appendix
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appendix D | Reagents, Instruments,