Background: Research into gene expression enables scientists to decipher the complex regulatory networks that control fundamental biological processes. Quantitative real-time PCR (qPCR) is a powerful and ubiquitous method for interrogation of gene expression. Accurate quantification is essential for correct interpretation of qPCR data. However, conventional relative and absolute quantification methodologies often give erroneous results or are laborious to perform. To overcome these failings, we developed an accurate, simple to use, universal calibrator, AccuCal.
Results: Herein, we show that AccuCal quantification can be used with either dye- or probe-based detection methods and is accurate over a dynamic range of ≥10(5) copies, for amplicons up to 500 base pairs (bp). By providing absolute quantification of all genes of interest, AccuCal exposes, and circumvents, the well-known biases of qPCR, thus allowing objective experimental conclusions to be drawn.
Conclusion: We propose that AccuCal supersedes the traditional quantification methods of PCR.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4784296 | PMC |
| http://dx.doi.org/10.1186/s12896-016-0256-y | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Sequence analysis of the 5' region of the chymotrypsin C (CTRC) gene in chronic pancreatitis.
Pancreatology
January 2025
Center for Gastroenterology, Department of Medicine, Albert Szent-Györgyi Medical School, University of Szeged, Szeged, Hungary; Hungarian Centre of Excellence for Molecular Medicine - University of Szeged, Translational Pancreatology Research Group, Szeged, Hungary. Electronic address:
Background/objectives: Loss-of-function chymotrypsin C (CTRC) variants increase the risk for chronic pancreatitis (CP) by reducing protective pancreatic CTRC activity. Variants in the 5' upstream region that includes the promoter might affect CTRC expression but have not been investigated to date. The aim of the present study was to address this knowledge gap.
View Article and Find Full Text PDFPersonalized treatment approaches in hepatocellular carcinoma.
Arab J Gastroenterol
January 2025
Endemic Medicine Department, Faculty of Medicine, Helwan University, Cairo, Egypt; Liver Disease Research Center, College of Medicine, King Saud University, Riyadh 11411, Saudi Arabia. Electronic address:
Personalized medicine is an emerging field that provides novel approaches to disease's early diagnosis, prevention, treatment, and prognosis based on the patient's criteria in gene expression, environmental factors, lifestyle, and diet. To date, hepatocellular carcinoma (HCC) is a significant global health burden, with an increasing incidence and significant death rates, despite advancements in surveillance, diagnosis, and therapeutic approaches. The majority of HCC lesions develop in patients with liver cirrhosis, carrying the risks of mortality associated with both the tumor burden and the cirrhosis.
View Article and Find Full Text PDFReconciliation of wheat 660 K and 90 K SNP arrays and their utilization in dough rheological properties of bread wheat.
J Adv Res
January 2025
Agronomy College / National Key Laboratory of Wheat and Maize Crop Science, Henan Agricultural University, Zhengzhou 450046 China. Electronic address:
Introduction: High-density Wheat 660 K and 90 K SNP arrays are powerful tools for understanding the genetic basis of wheat traits. However, their inconsistantly physical positions that were caused by different versions of Chinese Spring genome during developing arrays are confused and inconvenient for further application.
Objective: With the repid development of wheat geonome sequencing, we aim to reconciliate Wheat 660 K and 90 K SNP arrays in modern cultivar and reveal the genetic basis of dough rheological properties in bread wheat.
Phylogenetic analysis and homology modelling of a new Cry8A crystal protein expressed in a sporulating soil bacterium.
J Struct Biol
January 2025
Postgraduate Program in Industrial Biotechnology, Tiradentes University, Aracaju, Sergipe, Brazil; Department of Morphology, Federal University of Sergipe, São Cristóvão, Sergipe, Brazil. Electronic address:
Cry proteins, commonly found in gram-positive soil bacteria, are used worldwide as aerial sprays or in transgenic plants for controlling crop pest populations and as insect vectors. Via PCR analysis, a spore producing soil isolate (BV5) was speculated to encode a Cry gene. Partial nucleotide sequence of the amplified PCR fragment showed homology with the Cry8 genes present in GenBank.
View Article and Find Full Text PDFMatrix-mediated activation of murine fibroblast-like synoviocytes.
Exp Cell Res
January 2025
Translational Matrix Biology, University of Cologne, Medical Faculty, Cologne, Germany. Electronic address:
Fibroblast-like synoviocytes (FLS) are key cells promoting cartilage damage and bone loss in rheumatoid arthritis (RA). They are activated to assume an invasive and migratory phenotype. While mechanisms of FLS activation are unknown, evidence suggests that pre-damaged extracellular matrix (ECM) of the cartilage can trigger FLS activation.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!