کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1994518 | 1064790 | 2007 | 7 صفحه PDF | دانلود رایگان |
As key regulators of post-transcriptional gene expression, it is important to monitor the expression of microRNAs (miRNA) in diverse physiological and pathophysiological processes. Here, we describe a method for sensitive and accurate microarray-based expression profiling of miRNAs. The protocol focuses on the use of locked nucleic acid (LNA)-modified capture probes. LNAs are bicyclic nucleotide analogues that significantly increase the melting temperature (Tm) of hybrids with miRNAs. Mixed LNA/DNA capture probes thus can be designed for equal Tms for all miRNAs, which naturally cover a range between 45 and 74 °C. The protocols established are easy to apply, as they do not require RNA size selection and/or amplification of miRNAs. Moreover, they enable high affinity hybridizations yielding accurate signals that discriminate between single nucleotide differences and hence closely related miRNA family members.
Journal: Methods - Volume 43, Issue 2, October 2007, Pages 146–152