A novel multiplex qPCR method for assessing the comparative lengths of telomeres

Background: The comparative size of telomeres is taken into account to be associated to ailments corresponding to most cancers, growing older, and cardiovascular ailments. qPCR is presently one of many primary strategies for detecting telomere size. Nevertheless, because of the distinctive sequence of telomeres (extremely repetitive six-base sequence), it’s tough to design primers and probes to…

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computablegenomix

Ultra-sensitive AAV capsid detection by immunocapture-based qPCR following factor VIII gene transfer

Adeno-associated virus (AAV)-based gene remedy vectors are replication-incompetent and thus pose minimal threat for horizontal transmission or launch into the surroundings. In research with AAV5-FVIII-SQ (valoctocogene roxaparvovec), an investigational gene remedy for hemophilia A, residual vector DNA was detectable in blood, secreta, and excreta, nevertheless it remained unclear how lengthy structurally intact AAV5 vector capsids…

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computablegenomix

RT-qPCR Diagnostics: The “Drosten” SARS-CoV-2 Assay Paradigm

The reverse transcription quantitative polymerase chain reaction (RT-qPCR) is an established tool for the diagnosis of RNA pathogens. Its potential for automation has caused it to be used as a presence/absence diagnostic tool even when RNA quantification is not required. This technology has been pushed to the forefront of public awareness by the COVID-19 pandemic,…

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computablegenomix

A qPCR Assay for the Fast Detection and Quantification of Colletotrichum lupini

White lupin (Lupinus albus) represents an vital legume crop in Europe and different elements of the world on account of its excessive protein content material and potential for low-input agriculture. Nonetheless, most cultivars are vulnerable to anthracnose attributable to Colletotrichum lupini, a seed- and air-borne fungal pathogen that causes extreme yield losses. The goal of this work…

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computablegenomix

Validation of the analytical performance of nine commercial RT-qPCR kits for SARS-CoV-2 detection using certified reference material

The continued coronavirus illness 2019 (COVID-19) pandemic has turn into a public well being emergency. Though many reverse-transcription PCR (RT-PCR) assays have been developed, their efficiency, particularly sensitivity evaluation, has been insufficiently examined. On this examine, a preliminary comparability of the analytical sensitivity of 9 RT-qPCR kits from completely different producers was first performed utilizing…

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computablegenomix

What’s the norm in normalization? A frightening note on the use of RT-qPCR in the livestock science

Reverse-Transcription quantitative PCR (RT-qPCR) offers a precious device to check gene expression with beautiful sensitivity. To retain its inferential energy, user-introduced technical variability have to be decreased and accounted for. Deciding on a set of stably expressed inside management genes (ICG), validated for every experimental situation/pattern set, is broadly accepted as a dependable approach to…

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computablegenomix

Development of real-time RT-qPCR assays for the typing of two novel bluetongue virus genotypes derived from sheeppox vaccine

Beforehand, we reported the detection of two novel bluetongue virus (BTV) strains (SPvvvv/02 and SPvvvv/03), presumably representing new BTV genotypes, in a batch of sheeppox vaccine. We developed type-specific RT-qPCR assays (concentrating on genome section 2) for these two new BTV strains. The restrict of detection of each assays was 10 genome copies/μl and no…

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computablegenomix

Identification and validation of reference genes for RT-qPCR analysis in fetal rat pancreas

The selection of reference gene is essential for quantitative reverse transcriptase-polymerase chain response (RT-qPCR) assay. To display screen and decide the appropriate reference genes in fetal rat pancreas, we chosen eight candidate reference genes (Gapdh, Actb, Rn18 s, B2m, Rpl13a, Tbp, Ywhaz and Ubc), and evaluated the fidelity of gene expression from fetal rat pancreases…

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computablegenomix

An optimized stepwise algorithm combining rapid antigen and RT-qPCR for screening of COVID-19 patients

Background & intention: We investigated the mixture of speedy antigen detection (RAD) and RT-qPCR assays in a stepwise process to optimize the detection of COVID-19. Strategies: From August 2020 to November 2020, 43,399 sufferers had been screened in our laboratory for COVID-19 diagnostic by RT-qPCR utilizing nasopharyngeal swab. General, 4,691 of the 43,399 had been discovered to…

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computablegenomix

A New Specific and Sensitive RT-qPCR Method Based on Splinted 5′ Ligation for the Quantitative Detection of RNA Species Shorter than microRNAs

Not too long ago, we found a brand new household of unusually brief RNAs mapping to five.8S ribosomal RNA (rRNA) and which we named dodecaRNAs (doRNAs), based on the variety of core nucleotides (12 nt) their members include. To verify these small RNA-sequencing (RNA-Seq) knowledge, validate the existence of the 2 overly ample doRNAs-the minimal…

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