Comparative genomic hybridization (CGH) is a well-established technique in the field of molecular cytogenetics for performing the comprehensive analysis of chromosomal imbalances of entire human genomes. Genomic DNA from a tested specimen (test DNA) and a normal one (reference DNA) are differentially labeled and simultaneously hybridized to normal metaphase spreads. The ratio of fluorescence intensity along each normal chromosome is analyzed, allowing the detection of regions that are over- or underrepresented in the tested specimen. In this chapter, we describe the technique of CGH itself and a variant of it. The so-called micro-CGH (microdissection-based comparative genomic hybridization) technique allows molecular cytogenetics investigations of harvested and cytogenetically fixed (bone marrow) interphase nuclei. The latter must be microdissected from the surface of a coverslip, and the small amount of DNA present is amplified by a universal PCR using degenerate oligonucleotide primers (DOP-PCR). The PCR products are labeled and thus transformed into FISH probes. Micro-CGH has already been used in leukemia cytogenetics, as well as in clinical genetics.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Albertson DG, Pinkel D (2003) Genomic microarrays in human genetic disease and cancer. Hum Mol Genet 12 (Spec No 2):R145–R152
Chudoba I, Henn TK, Senger G, Claussen U (1997) ComParative genomic hybridization using DOP-PCR amplified DNA from small number of nuclei. Cs Pediatr 52:519–521
Daniely M, Aviram-Goldring A, Barkai G, Goldman B (1998) Detection of chromosomal aberration in fetuses arising from recurrent spontaneous abortion by comParative genomic hybridization. Hum Reprod 13:805–809
Gebhart E (2004) ComParative genomic hybridization (CGH): ten years of substantial progress in human solid tumor molecular cytogenetics. Cytogenet Genome Res 104:352–358
Gebhart E (2005) Genomic imbalances in human leukemia and lymphoma detected by comParative genomic hybridization. Int J Oncol 27:593–606
Gebhart E, Liehr T (2000) Patterns of genomic imbalances in human solid tumors (review). Int J Oncol 16:383–399
Heller A, Chudoba I, Bleck C, Senger G, Claussen U, Liehr T (2000a) Microdissection based comParative genomic hybridization analysis (micro-CGH) of secondary acute myelogenous leukemias. Int J Oncol 16:461–468
Heller A, Seidel J, Hübler A, Starke H, Beensen V, Senger G, Rocchi M, Wirth J, Chudoba I, Claussen U, Liehr T (2000b) Molecular cytogenetic characterisation of partial trisomy 9q in a case with pyloric stenosis and a review. J Med Genet 37:529–532
Heller A, Fricke HJ, Starke H, Loncarevic IF, Claussen U, Liehr T (2004) Characterization of a highly aberrant plasma cell leukemia karyotype: a case report. Oncol Rep 11:89–92
Kallioniemi A, Kallioniemi OP, Sudar D, Rutovitz D, Gray JW, Waldman F, Pinkel D (1992) ComParative genomic hybridization for molecular cytogenetic analysis of solid tumours. Science 258:818–821
Karst C, Heller A, Claussen U, Gebhart E, Liehr T (2005) Detection of cryptic chromosomal aberrations in the in vitro non-proliferating cells of acute myeloid leukemia. Int J Oncol 27:355–359
Kearney L, Horsley SW (2005) Molecular cytogenetics in haematological malignancy: current technology and future prospects. Chromosoma 114:286–294
Klijn C, Holstege H, de Ridder J, Liu X, Reinders M, Jonkers J, Wessels L (2008) Identification of cancer genes using a statistical framework for multiexperiment analysis of nondiscretized array CGH data. Nucleic Acids Res 36:e13
Knuutila S, Björkqvist AM, Autio K, Tarkkanen M, Wolf M, Monni O, Szymanska J, Larramendy ML, Tapper J, Pere H, El-Rifai W, Hemmer S, Wasenius VM, Vidgren V, Zhu Y (1998) DNA copy number amplifications in human neoplasms: review of comParative genomic hybridization studies. Am J Pathol 152:1107–1123
Paulsson K, Heidenblad M, Mörse H, Borg A, Fioretos T, Johansson B (2006) Identification of cryptic aberrations and characterization of translocation breakpoints using array CGH in high hyperdiploid childhood acute lymphoblastic leukemia. Leukemia 20:2002–2007
Sanlaville D, Baumann C, Lapierre JM, Romana S, Collot N, Cacheux V, Turleau C, Tachdjian G (1999) De novo inverted duplication 9p21pter involving telomeric repeated sequences. Am J Med Genet 83:125–131
Speicher MR, du Manoir S, Schröck E, Holtgreve-Grez H, Schoell B, Lengauer C, Cremer T, Ried T (1993) Molecular cytogenetic analysis of formalin-fixed, Paraffin-embedded solid tumors by comParative genomic hybridization after universal DNA-amplification. Hum Mol Genet 2:1907–1914
Starke H, Raida M, Trifonov V, Clement JH, Loncarevic IF, Heller A, Bleck C, Nietzel A, Rubtsov N, Claussen U, Liehr T (2001) Molecular cytogenetic characterization of an acquired minute supernumerary marker chromosome as the sole abnormality in a case clinically diagnosed as atypical Philadelphia-negative chronic myelogenous leukaemia. Br J Haematol 113:435–438
van Beers EH, Nederlof PM (2006) Array-CGH and breast cancer. Breast Cancer Res 8:210
Acknowledgments
Supported in part by Ernst-Abbe-Stiftung, INTAS (AISbl 03-51-4060), Deutsche Krebshilfe/Mildred Scheel Stiftung für Krebsforschung (70-3125-Li1), the IZKF Jena (Start-up S16, TP 3.7), TMWFK (B307-04004), DFG (436 ARM 17/5/06), Stiftung Leukämie, and Stefan-Morsch-Stiftung.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Springer-Verlag Berlin Heidelberg
About this protocol
Cite this protocol
Heller, A., Liehr, T. (2009). Micro-CGH: Microdissection-Based Comparative Genomic Hybridization. In: Liehr, T. (eds) Fluorescence In Situ Hybridization (FISH) — Application Guide. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-70581-9_34
Download citation
DOI: https://doi.org/10.1007/978-3-540-70581-9_34
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-70580-2
Online ISBN: 978-3-540-70581-9
eBook Packages: Springer Protocols