Next Article in Journal
Quantitative Trait Locus and Brain Expression of HLA-DPA1 Offers Evidence of Shared Immune Alterations in Psychiatric Disorders
Previous Article in Journal
Acknowledgement to Reviewers of Microarrays in 2015
Article Menu

Export Article

From the third issue of 2017, Microarrays has changed its name to High-Throughput.

Open AccessArticle
Microarrays 2016, 5(1), 5; doi:10.3390/microarrays5010005

A Double-Hybridization Approach for the Transcription- and Amplification-Free Detection of Specific mRNA on a Microarray

Center for Advanced Bioanalysis GmbH, Gruberstrasse 40-42, 4020 Linz, Austria
Department of Molecular Biology, University of Salzburg, 5020 Salzburg, Austria
Author to whom correspondence should be addressed.
Academic Editor: Massimo Negrini
Received: 17 December 2015 / Revised: 5 February 2016 / Accepted: 15 February 2016 / Published: 23 February 2016
View Full-Text   |   Download PDF [1464 KB, uploaded 23 February 2016]   |  


A double-hybridization approach was developed for the enzyme-free detection of specific mRNA of a housekeeping gene. Targeted mRNA was immobilized by hybridization to complementary DNA capture probes spotted onto a microarray. A second hybridization step of Cy5-conjugated label DNA to another section of the mRNA enabled specific labeling of the target. Thus, enzymatic artifacts could be avoided by omitting transcription and amplification steps. This manuscript describes the development of capture probe molecules used in the transcription- and amplification-free analysis of RPLP0 mRNA in isolated total RNA. An increase in specific signal was found with increasing length of the target-specific section of capture probes. Unspecific signal comprising spot autofluorescence and unspecific label binding did not correlate with the capture length. An additional spacer between the specific part of the capture probe and the substrate attachment site increased the signal significantly only on a short capture probe of approximately 30 nt length. View Full-Text
Keywords: microarray; mRNA detection; enzyme-free; gene expression; fluorescence microscopy microarray; mRNA detection; enzyme-free; gene expression; fluorescence microscopy

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Haider, M.; Haselgrübler, T.; Sonnleitner, A.; Aberger, F.; Hesse, J. A Double-Hybridization Approach for the Transcription- and Amplification-Free Detection of Specific mRNA on a Microarray. Microarrays 2016, 5, 5.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Article Access Statistics



[Return to top]
Microarrays EISSN 2076-3905 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top