Human and mouse oligonucleotide-based array CGH

被引:55
|
作者
van den IJssel, P
Tijssen, M
Chin, SF
Eijk, P
Carvalho, B
Hopmans, E
Holstege, H
Bangarusamy, DK
Jonkers, J
Meijer, GA
Caldas, C
Ylstra, B
机构
[1] Vrije Univ Amsterdam, Med Ctr, Microarray Core Facil, Dept Pathol, NL-1087 BT Amsterdam, Netherlands
[2] Univ Cambridge, Dept Oncol, Canc Genom Program, Cambridge, England
[3] Netherlands Canc Inst, Div Mol Biol, NL-1066 CX Amsterdam, Netherlands
[4] Inst Biomed Sci, Genome Inst Singapore, Singapore, Singapore
关键词
D O I
10.1093/nar/gni191
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Array-based comparative genomic hybridization is a high resolution method for measuring chromosomal copy number changes. Here we present a validated protocol using in-house spotted oligonucleotide libraries for array comparative genomic hybridization (CGH). This oligo array CGH platform yields reproducible results and is capable of detecting single copy gains, multi-copy amplifications as well as homozygous and heterozygous deletions as small as 100 kb with high resolution. A human oligonucleotide library was printed on amine binding slides. Arrays were hybridized using a hybstation and analysed using BlueFuse feature extraction software, with > 95% of spots passing quality control. The protocol allows as little as 300 ng of input DNA and a 90% reduction of Cot-1 DNA without compromising quality. High quality results have also been obtained with DNA from archival tissue. Finally, in addition to human oligo arrays, we have applied the protocol successfully to mouse oligo arrays. We believe that this oligo-based platform using 'off-the-shelf' oligo libraries provides an easy accessible alternative to BAC arrays for CGH, which is cost-effective, available at high resolution and easily implemented for any sequenced organism without compromising the quality of the results.
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页数:9
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