---
OA_type: closed access
_id: '4145'
abstract:
- lang: eng
text: The detection of microRNAs (miRNAs) at single-cell resolution is important
for studying the role of these posttranscriptional regulators. Here, we use a
dual-fluorescent green fluorescent protein (GFP)-reporter/monomeric red fluorescent
protein (mRFP)-sensor (DFRS) plasmid, injected into zebrafish blastomeres or electroporated
into defined tissues of mouse embryos in utero or ex utero, to monitor the dynamics
of specific miRNAs in individual live cells. This approach reveals, for example,
that in the developing mouse central nervous system,, miR-124a is expressed not
only in postmitotic neurons but also in neuronal progenitor cells. Collectively,
our results demonstrate that acute administration of DFRS plasmids.offers an alternative
to previous in situ hybridization and transgenic approaches and allows the monitoring
of miRNA appearance and disappearance in defined cell lineages during vertebrate
development.
article_processing_charge: No
article_type: original
author:
- first_name: Davide
full_name: Tonelli, Davide
last_name: Tonelli
- first_name: Frederico
full_name: Calegari, Frederico
last_name: Calegari
- first_name: Ji
full_name: Fei, Ji
last_name: Fei
- first_name: Tadashi
full_name: Nomura, Tadashi
last_name: Nomura
- first_name: Noriko
full_name: Osumi, Noriko
last_name: Osumi
- first_name: Carl-Philipp J
full_name: Heisenberg, Carl-Philipp J
id: 39427864-F248-11E8-B48F-1D18A9856A87
last_name: Heisenberg
orcid: 0000-0002-0912-4566
- first_name: Wieland
full_name: Huttner, Wieland
last_name: Huttner
citation:
ama: Tonelli D, Calegari F, Fei J, et al. Single-cell detection of microRNAs in
developing vertebrate embryos after acute administration of a dual-fluorescence
reporter/sensor plasmid. Biotechniques. 2006;41(6):727-732. doi:10.2144/000112296
apa: Tonelli, D., Calegari, F., Fei, J., Nomura, T., Osumi, N., Heisenberg, C.-P.
J., & Huttner, W. (2006). Single-cell detection of microRNAs in developing
vertebrate embryos after acute administration of a dual-fluorescence reporter/sensor
plasmid. Biotechniques. Informa Healthcare. https://doi.org/10.2144/000112296
chicago: Tonelli, Davide, Frederico Calegari, Ji Fei, Tadashi Nomura, Noriko Osumi,
Carl-Philipp J Heisenberg, and Wieland Huttner. “Single-Cell Detection of MicroRNAs
in Developing Vertebrate Embryos after Acute Administration of a Dual-Fluorescence
Reporter/Sensor Plasmid.” Biotechniques. Informa Healthcare, 2006. https://doi.org/10.2144/000112296.
ieee: D. Tonelli et al., “Single-cell detection of microRNAs in developing
vertebrate embryos after acute administration of a dual-fluorescence reporter/sensor
plasmid,” Biotechniques, vol. 41, no. 6. Informa Healthcare, pp. 727–732,
2006.
ista: Tonelli D, Calegari F, Fei J, Nomura T, Osumi N, Heisenberg C-PJ, Huttner
W. 2006. Single-cell detection of microRNAs in developing vertebrate embryos after
acute administration of a dual-fluorescence reporter/sensor plasmid. Biotechniques.
41(6), 727–732.
mla: Tonelli, Davide, et al. “Single-Cell Detection of MicroRNAs in Developing Vertebrate
Embryos after Acute Administration of a Dual-Fluorescence Reporter/Sensor Plasmid.”
Biotechniques, vol. 41, no. 6, Informa Healthcare, 2006, pp. 727–32, doi:10.2144/000112296.
short: D. Tonelli, F. Calegari, J. Fei, T. Nomura, N. Osumi, C.-P.J. Heisenberg,
W. Huttner, Biotechniques 41 (2006) 727–732.
date_created: 2018-12-11T12:07:12Z
date_published: 2006-12-01T00:00:00Z
date_updated: 2026-08-26T09:09:45Z
day: '01'
doi: 10.2144/000112296
extern: '1'
external_id:
pmid:
- '17191618 '
fulldoi: https://doi.org/10.2144/000112296
intvolume: ' 41'
issue: '6'
language:
- iso: eng
month: '12'
oa_version: None
page: 727 - 732
pmid: 1
publication: Biotechniques
publication_identifier:
eissn:
- 1940-9818
issn:
- 0736-6205
publication_status: published
publisher: Informa Healthcare
publist_id: '1974'
status: public
title: Single-cell detection of microRNAs in developing vertebrate embryos after acute
administration of a dual-fluorescence reporter/sensor plasmid
type: journal_article
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 41
year: '2006'
...
---
_id: '3422'
abstract:
- lang: eng
text: Quantitative real-time PCR represents a highly sensitive and powerful technique
for the quantitation of nucleic acids. It has a tremendous potential for the high-throughput
analysis of gene expression in research and routine diagnostics. However, the
major hurdle is not the practical performance of the experiments themselves but
rather the efficient evaluation and the mathematical and statistical analysis
of the enormous amount of data gained by this technology, as these functions are
not included in the software provided by the manufacturers of the detection systems.
In this work, we focus on the mathematical evaluation and analysis of the data
generated by quantitative real-time PCR, the calculation of the final results,
the propagation of experimental variation of the measured values to the final
results, and the statistical analysis. We developed a Microsoft Excel-based software
application coded in Visual Basic for Applications, called Q-Gene, which addresses
these points. Q-Gene manages and expedites the planning, performance, and evaluation
of quantitative real-time PCR experiments, as well as the mathematical and statistical
analysis, storage, and graphical presentation of the data. The Q-Gene software
application is a tool to cope with complex quantitative real-time PCR experiments
at a high-throughput scale and considerably expedites and rationalizes the experimental
setup, data analysis, and data management while ensuring highest reproducibility.
article_processing_charge: No
article_type: original
author:
- first_name: Patrick
full_name: Müller, Patrick
last_name: Müller
- first_name: Harald L
full_name: Janovjak, Harald L
id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
last_name: Janovjak
orcid: 0000-0002-8023-9315
- first_name: Andre
full_name: Miserez, Andre
last_name: Miserez
- first_name: Zuzana
full_name: Dobbie, Zuzana
last_name: Dobbie
citation:
ama: Müller P, Janovjak HL, Miserez A, Dobbie Z. Processing of gene expression data
generated by quantitative real-time RT-PCR. Biotechniques. 2002;32(6):1372-1379.
apa: Müller, P., Janovjak, H. L., Miserez, A., & Dobbie, Z. (2002). Processing
of gene expression data generated by quantitative real-time RT-PCR. Biotechniques.
Informa Healthcare.
chicago: Müller, Patrick, Harald L Janovjak, Andre Miserez, and Zuzana Dobbie. “Processing
of Gene Expression Data Generated by Quantitative Real-Time RT-PCR.” Biotechniques.
Informa Healthcare, 2002.
ieee: P. Müller, H. L. Janovjak, A. Miserez, and Z. Dobbie, “Processing of gene
expression data generated by quantitative real-time RT-PCR,” Biotechniques,
vol. 32, no. 6. Informa Healthcare, pp. 1372–1379, 2002.
ista: Müller P, Janovjak HL, Miserez A, Dobbie Z. 2002. Processing of gene expression
data generated by quantitative real-time RT-PCR. Biotechniques. 32(6), 1372–1379.
mla: Müller, Patrick, et al. “Processing of Gene Expression Data Generated by Quantitative
Real-Time RT-PCR.” Biotechniques, vol. 32, no. 6, Informa Healthcare, 2002,
pp. 1372–79.
short: P. Müller, H.L. Janovjak, A. Miserez, Z. Dobbie, Biotechniques 32 (2002)
1372–1379.
date_created: 2018-12-11T12:03:15Z
date_published: 2002-06-01T00:00:00Z
date_updated: 2023-07-17T11:29:06Z
day: '01'
extern: '1'
external_id:
pmid:
- '12074169'
intvolume: ' 32'
issue: '6'
language:
- iso: eng
month: '06'
oa_version: None
page: 1372 - 1379
pmid: 1
publication: Biotechniques
publication_identifier:
issn:
- 0736-6205
publication_status: published
publisher: Informa Healthcare
publist_id: '2979'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Processing of gene expression data generated by quantitative real-time RT-PCR
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 32
year: '2002'
...