Measuring differential gene expression by short read sequencing: quantitative comparison to 2-channel gene expression microarrays

被引:127
作者
Bloom, Joshua S. [2 ]
Khan, Zia [3 ]
Kruglyak, Leonid [1 ,4 ,5 ]
Singh, Mona [1 ,3 ]
Caudy, Amy A. [1 ]
机构
[1] Princeton Univ, Lewis Sigler Inst Integrat Genom, Princeton, NJ 08544 USA
[2] Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA
[3] Princeton Univ, Dept Comp Sci, Princeton, NJ 08544 USA
[4] Princeton Univ, Dept Ecol & Evolutionary Biol, Princeton, NJ 08544 USA
[5] Princeton Univ, Howard Hughes Med Inst, Princeton, NJ 08544 USA
来源
BMC GENOMICS | 2009年 / 10卷
基金
美国国家卫生研究院;
关键词
RNA-SEQ; TRANSCRIPTOME; YEAST; MPSS; ARABIDOPSIS; RESOLUTION; GENOME; ARRAYS; PCR;
D O I
10.1186/1471-2164-10-221
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: High-throughput cDNA synthesis and sequencing of poly(A)-enriched RNA is rapidly emerging as a technology competing to replace microarrays as a quantitative platform for measuring gene expression. Results: Consequently, we compared full length cDNA sequencing to 2-channel gene expression microarrays in the context of measuring differential gene expression. Because of its comparable cost to a gene expression microarray, our study focused on the data obtainable from a single lane of an Illumina 1 G sequencer. We compared sequencing data to a highly replicated microarray experiment profiling two divergent strains of S. cerevisiae. Conclusion: Using a large number of quantitative PCR (qPCR) assays, more than previous studies, we found that neither technology is decisively better at measuring differential gene expression. Further, we report sequencing results from a diploid hybrid of two strains of S. cerevisiae that indicate full length cDNA sequencing can discover heterozygosity and measure quantitative allele-specific expression simultaneously.
引用
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页数:10
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