7500 real time polymerase chain reaction system  (Thermo Fisher)

Journal: Neuroendocrinology

Article Title: Brain-Derived Neurotrophic Factor and Supraoptic Vasopressin Neurons in Hyponatremia

doi: 10.1159/000503723

Figure Lengend Snippet: a Representative digital image of mCherry fluorescence in the SON illustrating a successful injection. b Quantitative real-time PCR data showing BDNF mRNA expression from the SON of rats with successful SON injections. c AVP hnRNA expression from the SONs of rats with successful SON injections. Groups: Sham-ligated rats injected with SCR virus (Sham SCR, n = 5); Sham rats injected with ­shBDNF virus (Sham shBDNF, n = 6); BDL rats injected with SCR virus (BDL SCR, n = 6), and BDL rats injected with shBDNF virus (BDL shBDNF, n = 6). Data are mean ± SEM. * p

Article Snippet: The SONs collected from LCM were used to measure changes in BDNF mRNA and AVP hnRNA using quantitative real-time PCR.

Techniques: Fluorescence, Injection, Real-time Polymerase Chain Reaction, Expressing

Journal: Nature Communications

Article Title: Performance of the Trioplex real-time RT-PCR assay for detection of Zika, dengue, and chikungunya viruses

doi: 10.1038/s41467-018-03772-1

Figure Lengend Snippet: Analytical performance comparison between Trioplex assay multiplex and the ZIKV singleplex format assay using small volume and large volume RNA extraction. Normal human serum or urine was contrived with ZIKV at a dilution of 1:10 before the limit of detection (1:10 BLoD), at the limit of detection (LoD), and at 1:10 after the limit of detection (1:10 ALoD). Twenty replicates of every dilution were extracted using the MagNA Pure 96 instrument (Roche) and tested by Trioplex assay multiplex or ZIKV singleplex format assay on the ABI 7500 Fast Dx or the QuantStudio Dx instruments. a Compares the mean genome copy equivalents per PCR reaction (GCE/rxn) of viral RNA extracted from serum at each dilution on the ABI 7500 Fast Dx instrument. A linear regression was plotted for multiplex with small volume protocol (Sv) (0.2 mL) (black straight line), singleplex assay with small volume protocol (gray straight line), multiplex with large volume protocol (Lv) (1 mL) (black dashed line). and singleplex assay with large volume protocol (gray dashed line). b Compares the mean genome copy equivalents per PCR reaction (GCE/rxn) of viral RNA extracted from serum or urine at each dilution on the QuantStudio Dx instrument. A linear regression was plotted for multiplex with small volume protocol serum (Sv) (black straight line), multiplex with large volume protocol serum (Lv) (gray straight line), multiplex with small volume protocol urine (Lv) (black dashed line), and multiplex with large volume protocol urine (gray dashed line). Error bars represent GCE/mL standard deviation. The CT values for every dilution replicate in serum tested was plotted for c small volume and d large volume extractions

Article Snippet: The fluorophore-labeled hydrolysis probes bind to the amplified DNA target fragment and the intensity of the fluorescent signal is captured by a real-time PCR instrument: (ABI 7500 fast Dx (ThermoFisher) or QuantStudio Dx (ThermoFisher)).

Techniques: Multiplex Assay, RNA Extraction, Polymerase Chain Reaction, Singleplex Assay, Standard Deviation

Journal: Frontiers in Immunology

Article Title: MicroRNA-130a Contributes to Type-2 Classical DC-activation in Sjögren's Syndrome by Targeting Mitogen- and Stress-Activated Protein Kinase-1

doi: 10.3389/fimmu.2019.01335

Figure Lengend Snippet: MSK1 inhibition reduces cDC2 activation and inhibits the production of pro-inflammatory cytokines. Isolated cDC2s from buffy coats were treated with H89 (10 μM), SB 747651A (SB; 10 μM), Ro 31-8220 (Ro; 5 μM), or left untreated for 1 h. Then, except for medium control, cells were stimulated with TLR4L (100 ng/mL) for 6 h. FACS was used to assess cell-viability (A) and the expression of CD80 and CD83 shown as median fluorescence intensity (MFI) (B) . Cytokine production upon TLR4L stimulation in the presence or absence of MSK1 inhibitors was measured by qRT-PCR (C) and ELISA (D) . Results are represented as median ± IQR. * p

Article Snippet: Detailed descriptions of stable isotope labeling of amino acids in cell culture (SILAC), selection of in silico predicted miRNA targets, quantitative real-time PCR and cytokine analysis are provided in the Online Supplementary Methods.

Techniques: Inhibition, Activation Assay, Isolation, FACS, Expressing, Fluorescence, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Performance of the Trioplex real-time RT-PCR assay for detection of Zika, dengue, and chikungunya viruses

doi: 10.1038/s41467-018-03772-1

Figure Lengend Snippet: Clinical performance of the Trioplex assay across specimen types. Clinical specimens collected concurrently from 373 cases with previous Zika determination in the acute stage were tested. RNA was extracted with the MagNA Pure 96 small volume external lysis protocol from 373 case-paired serum, 373 urine, and 345 whole blood-EDTA specimens and tested with the Trioplex assay in multiplex format in the ABI 7500 Fast Dx instrument. a Correlation of CT values between case-matching serum and urine specimens; R 2 = 0.36 p

Article Snippet: Performance of the Trioplex assay was evaluated using RNA extractions from the MagNA Pure LC 2.0, MagNA Pure 96, MagNA Pure Compact, and NucliSENS® easyMag® platforms and run on the ABI 7500 Fast Dx and QuantStudio Dx real-time PCR instruments.

Techniques: Lysis, Multiplex Assay