PUMA publications for /tag/standard;%20expression%7D%20myown%20qrt-pcr;https://puma.ub.uni-stuttgart.de/tag/standard;%20expression%7D%20myown%20qrt-pcr;PUMA RSS feed for /tag/standard;%20expression%7D%20myown%20qrt-pcr;2024-03-29T16:58:41+01:00Quantifying cytosolic messenger RNA concentrations in Escherichia coli
using real-time polymerase chain reaction for a systems biology approachhttps://puma.ub.uni-stuttgart.de/bibtex/22af41d00232e2b7ff14070cdc783d6e8/siemannherzbergsiemannherzberg2018-01-25T13:38:08+01:00Absolute Gene Internal concentrations; expression} mRNA myown qRT-PCR; quantification; standard; {Cytosolic <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tom Schuhmacher" itemprop="url" href="/person/184c2f964d514c704ed8241b609338635/author/0"><span itemprop="name">T. Schuhmacher</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Karin Lemuth" itemprop="url" href="/person/184c2f964d514c704ed8241b609338635/author/1"><span itemprop="name">K. Lemuth</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Timo Hardiman" itemprop="url" href="/person/184c2f964d514c704ed8241b609338635/author/2"><span itemprop="name">T. Hardiman</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Gabi Vacun" itemprop="url" href="/person/184c2f964d514c704ed8241b609338635/author/3"><span itemprop="name">G. Vacun</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Matthias Reuss" itemprop="url" href="/person/184c2f964d514c704ed8241b609338635/author/4"><span itemprop="name">M. Reuss</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Martin Siemann-Herzberg" itemprop="url" href="/person/184c2f964d514c704ed8241b609338635/author/5"><span itemprop="name">M. Siemann-Herzberg</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">ANALYTICAL BIOCHEMISTRY</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">398 </span></span>(<span itemprop="issueNumber">2</span>):
<span itemprop="pagination">212-217</span></em> </span>(<em><span>March 2010<meta content="March 2010" itemprop="datePublished"/></span></em>)</span>Thu Jan 25 13:38:08 CET 2018{525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA}{ANALYTICAL BIOCHEMISTRY}{MAR 15}{2}{212-217}{Quantifying cytosolic messenger RNA concentrations in Escherichia coli
using real-time polymerase chain reaction for a systems biology approach}{Article}{398}{2010}Absolute Gene Internal concentrations; expression} mRNA myown qRT-PCR; quantification; standard; {Cytosolic {Current messenger RNA (mRNA) quantification methods are sophisticated
tools for the analysis of gene regulation. However, these methods are
not suitable for more complex quantitative approaches such as the
mathematical modeling of the in vivo regulation of transcription where
dynamic cytosolic mRNA concentrations need to be taken into
consideration. In the current study, the ``standard curve method{''} for
quantitative reverse transcription real-time polymerase chain reaction
(qRT-PCR) was extended by including an internal RNA standard. This
standard enables transcript losses that occur during the process, as
well as variations resulting from nonquantitative processes, to be
accounted for. The use of an internal standard yielded transcript
concentration estimates that were on average seven times higher than
those in cases where an internal standard is omitted. Choosing the cra
modulon in Escherichia coli as an example, the method applied shows that
the regulation of the Cra protein, as well as the growth rate-dependent
regulation, need to be taken into consideration. The new method, which
enables the determination of cytosolic mRNA concentrations, allows the
quantitative representation of transcriptional dynamics. This is an
important aspect of the analysis of the complex interactions of
metabolism and regulation and in the application of mathematical
modeling for systems biology. (C) 2009 Elsevier Inc. All rights
reserved.}