Ekstraksi Dna Dan Amplifikasi Its Rdna Isolat Fungi Endofit Lbkurcc67 Umbi Tanaman Dahlia (Dahlia Variabilis)

> Endophyte fungi lives in the plant tissues without causing harm to their host and has known that can produce secondary metabolite and extracellular enzyme. Fungi LBKURCC67 is an endophyte fungi that was isolated from tubers of yellow flowered Dahlia variabilis in Padang Panjang, West Sumatera. Species of fungi LBKURCC67 isolate was not known exactly because the morphology identification has been previously is appropriate for genus level. The accurate identification to known species is molecular with phylogenetic analysis. Before determine species in molecular analysis, it must extraction and amplification DNA. This research aims to optimation of DNA extraction and rDNA amplification in Internal Transcribed Spacer (ITS) regions with Polymerase Chain Reactions (PCR) methods. The DNA of fungi LBKURCC67 isolate was extract with Wizard Genomic DNA Purification kit ex Promega Corp. (Madison, USA) from cultur mycelium. The result shows extraction DNA was success from fourth days mycelia. Optimum condition for rDNA amplification with PCR were used ITS5 and ITS4 primers and 41°C annealing temperature. Electrophoresis analysis shows molecular weight of DNA isolate is 16.951 bp and molecular weight of PCR product is 583 bp

CSMG neurons express mu opioid (A) and free fatty acid (B) receptors.

<p>A, left, superimposed Ca²⁺ current traces (evoked as described for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125566#pone.0125566.g003" target="_blank">Fig 3</a>) in the absence (lower traces, black) and presence (upper traces, gray) of 10 μM DAMGO for sham control and septic rats. A, right, DAMGO concentration-response relationship of CSMG neurons from sham control (▲) and septic (●) rats. Each data point represents the mean (± SE) DAMGO-mediated prepulse current inhibition from 3 to 9 neurons, (except for 0.1 nM DAMGO for sham group where n = 2). Both smooth curves were obtained by fitting the points with the Hill equation and the fits were not significantly different (P > 0.05). The legend indicates the EC_<b>50</b> values (μM). B, left, propionate—mediated Ca²⁺ current inhibition in sham control and septic rats. B, right, summary plot indicates the mean (± SE) current inhibition produced by 1 and 3 mM propionate. Numbers in parenthesis indicate the number of neurons tested.</p

Normalized current-voltage (I-V) relationships of acutely isolated CSMG neurons from sham control (A) and septic (B) rats 48 hr post-sepsis induction by CLP.

<p>The I-V curves represent the mean Ca²⁺ current amplitude for each test potential. Ca²⁺ currents were evoked every 3 s with a 70 ms pulse from a holding potential of -80 mV to test potential between -60 and +60 mV. The current amplitude was measured 10 ms following the onset of the test pulse and normalized to +15 mV. The superimposed Ca²⁺ current traces shown to the left were evoked to potentials from -25 to +40 mV. The number of neurons tested for each group is shown in parenthesis.</p