Reference: Jin K, et al. (2011) Gene Expression Profiling via Multigene Concatemers. PLoS One 6(1):e15711

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Abstract


We established a novel method, Gene Expression Profiling via Multigene Concatemers (MgC-GEP), to study multigene expression patterns simultaneously. This method consists of the following steps: (1) cDNA was obtained using specific reverse primers containing an adaptor. (2) During the initial 1-3 cycles of polymerase chain reaction (PCR), the products containing universal adaptors with digestion sites at both termini were amplified using specific forward and reverse primers containing the adaptors. (3) In the subsequent 4-28 cycles, the universal adaptors were used as primers to yield products. (4) The products were digested and ligated to produce concatemers. (5) The concatemers were cloned into the vector and sequenced. Then, the occurrence of each gene tag was determined. To validate MgC-GEP, we analyzed 20 genes in Saccharomyces cerevisiae induced by weak acid using MgC-GEP combined with real-time reverse transcription (RT)-PCR. Compared with the results of real-time RT-PCR and the previous reports of microarray analysis, MgC-GEP can precisely determine the transcript levels of multigenes simultaneously. Importantly, MgC-GEP is a cost effective strategy that can be widely used in most laboratories without specific equipment. MgC-GEP is a potentially powerful tool for multigene expression profiling, particularly for moderate-throughput analysis.

Reference Type
Evaluation Study | Journal Article | Research Support, Non-U.S. Gov't
Authors
Jin K, Zheng X, Xia Y
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Gene Ontology Annotations


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Gene/Complex Qualifier Gene Ontology Term Aspect Annotation Extension Evidence Method Source Assigned On Reference

Phenotype Annotations


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Gene Phenotype Experiment Type Mutant Information Strain Background Chemical Details Reference

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Gene Disease Ontology Term Qualifier Evidence Method Source Assigned On Reference

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Post-translational Modifications


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Site Modification Modifier Reference

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

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Interactor Interactor Assay Annotation Action Modification Source Reference

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Gene Species Gene ID Strain background Direction Details Source Reference