qpcrbio probe mix hi rox  (PCR Biosystems Ltd)

Journal: Nucleic Acids Research

Article Title: The two-step purification method ViREn identifies a single NSUN6-mediated 5-methylcytosine modification promoting dengue virus RNA genome turnover

doi: 10.1093/nar/gkag003

Figure Lengend Snippet: Establishment of ViREn. ( A ) Schematics of the ViREn workflow. DENV RNA from either infected cells or virions was separated by sucrose gradient ultracentrifugation. DENV gRNA and sfRNA were further enriched using antisense affinity capture. The purified DENV gRNA was subjected to RNA sequencing-based approaches for the identification and mapping of m 5 C modifications. The distribution of RNAs extracted from infected cells ( B ) and virions ( C ) across sucrose density gradient fractions was analyzed by denaturing agarose gel and corresponding northern blot using a hybridization probe targeting DENV 3′UTR. The percentage of DENV gRNA in each fraction is shown at the bottom (mean ± SD, n = 3). ( D ) DENV gRNA collected in fraction 11 was further purified by affinity capture. Its integrity was visualized by northern blotting. ( E ) The level of DENV gRNA and 28S rRNA contaminant, before and after affinity capture, was measured by probe-based RT-qPCR (mean ± SD, n = 3). ( F ) The total RNA composition of fraction 11 before and after affinity capture was analyzed by Illumina-based RNA-seq ( n = 3). Shown is the proportion of reads in each sample corresponding to DENV gRNA, human genome, rRNAs, and unmapped reads (mean ± SD, n = 3).

Article Snippet: Briefly, 3 μl of extracted RNA were mixed with 12 μl qPCR mix (qPCRBIO Probe 1-Step Go Lo-ROX, PCR Biosystems) according to the manufacturer instructions and using the following cycling protocol: 50°C 10 min, 95°C 1 min, 95°C 10 s, 60°C 1 min; steps 3–4 were repeated 39 times.

Techniques: Infection, Purification, RNA Sequencing, Agarose Gel Electrophoresis, Northern Blot, Hybridization, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Morphological and molecular characterisation of Orientia tsutsugamushi grown in tick cells

doi: 10.64898/2025.12.15.694307

Figure Lengend Snippet: Orientia tsutsugamushi grows in tick cell lines. A) Graphs showing bacterial copy number per mL of the Ot strain TA686 at 25, 32, and 35 °C in ISE6 and BME/CTVM23 cell lines. B) Bacterial copy number of the Ot strain Karp in ISE6 and BME/CTVM23 at 32 °C. Bacterial load was measured by qPCR using primers against the conserved single-copy bacterial gene tsa47 .

Article Snippet: The qPCR mixture consisted of 1X qPCR Probe Mix LO-ROX (PCR Biosystems, UK), 0.1 μM of each forward and reverse primer, 0.2 μM probe, RT-PCR grade water (Thermo Scientific), and 1 μL of extracted DNA.

Techniques: