Abstract:
:The expression pattern and regulatory functions of microRNAs (miRNAs) are intensively investigated in various tissues, cell types and disorders. Differential miRNA expression signatures have been revealed in healthy and unhealthy tissues using high-throughput profiling methods. For further analyses of miRNA signatures in biological samples, we describe here a simple and efficient method to detect multiple miRNAs simultaneously in total RNA. The size-coded ligation-mediated polymerase chain reaction (SL-PCR) method is based on size-coded DNA probe hybridization in solution, followed-by ligation, PCR amplification and gel fractionation. The new method shows quantitative and specific detection of miRNAs. We profiled miRNAs of the let-7 family in a number of organisms, tissues and cell types and the results correspond with their incidence in the genome and reported expression levels. Finally, SL-PCR detected let-7 expression changes in human embryonic stem cells as they differentiate to neuron and also in young and aged mice brain and bone marrow. We conclude that the method can efficiently reveal miRNA signatures in a range of biological samples.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Arefian E,Kiani J,Soleimani M,Shariati SA,Aghaee-Bakhtiari SH,Atashi A,Gheisari Y,Ahmadbeigi N,Banaei-Moghaddam AM,Naderi M,Namvarasl N,Good L,Faridani ORdoi
10.1093/nar/gkr214subject
Has Abstractpub_date
2011-07-01 00:00:00pages
e80issue
12eissn
0305-1048issn
1362-4962pii
gkr214journal_volume
39pub_type
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