TRIzol and Alu qPCR-based quantification of metastatic seeding within the skeleton
نویسندگان
چکیده
Current methods for detecting disseminated tumor cells in the skeleton are limited by expense and technical complexity. We describe a simple and inexpensive method to quantify, with single cell sensitivity, human metastatic cancer in the mouse skeleton, concurrently with host gene expression, using TRIzol-based DNA/RNA extraction and Alu sequence qPCR amplification. This approach enables precise quantification of tumor cells and corresponding host gene expression during metastatic colonization in xenograft models.
منابع مشابه
Pitfalls of DNA Quantification Using DNA-Binding Fluorescent Dyes and Suggested Solutions
The Qubit fluorometer is a DNA quantification device based on the fluorescence intensity of fluorescent dye binding to double-stranded DNA (dsDNA). Qubit is generally considered useful for checking DNA quality before next-generation sequencing because it measures intact dsDNA. To examine the most accurate and suitable methods for quantifying DNA for quality assessment, we compared three quantif...
متن کاملMetastatic Tumors to Craniofacial Skeleton: Analysis of Two Cases and Review of the Literature
Craniofacial skeletal metastasis is a rare presentation of advanced prostate cancer. This is a report of a 69-year-old man who presented with numbness of the right lower lip and recently ill-fitting lower denture. Based on the medical history of benign prostate hyperplasia (BPH) and suspicion of a metastatic tumor, prostate core needle biopsy was performed. Histology of the prostate biopsy con...
متن کاملAbsolute quantification of murine interleukine-4, interleukine- 10 and interferon-γ gene transcripts using Real Time PCR
The study of cytokines gene expression is quite important in various conditions of health and disease for the evaluation of clinical responses to new vaccination approaches. An absolute quantification is based on a calibration curve and production of standard controls to achieve more reliable results than in relative system. In this study we attempted to construct standard controls to evaluate ...
متن کاملAlu DNA Polymorphism of Human Tissue Plasminogen Activator (tPA) Gene in Diabetic Jordanian Patients Patients
Background: Hypercoagulability and hypofibrinolysis are among the symptoms exhibited by diabetic patients. Our study aimed to address the polymorphic nature of Alu DNA fragment in the human tissue plasminogen activator gene within diabetes mellitus (DM) Jordanian patients. Methods: Genomic DNA was isolated from 76 DM patients and 60 non-diabetic Jordanian individuals, and the Alu fragment was a...
متن کاملTRIzol-based RNA Extraction: A Reliable Method for Gene Expression Studies
RNA extraction is a prerequisite technique for gene expression studies, analyzing the etiology and disease progression, treatment effects, as well as designing the diagnostic methods. Although many RNA extraction kits have been commercialized, but these kits are expensive and are not accessible in some countries. Many studies have shown that TRIzol is an applicable material for the RNA extracti...
متن کامل