Molecular Genetic TNF Alpha Gene Polymorphism - PCR
Test Code
MOG61
Also Known As
No data available
Test Parameters Included
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Department
MOLECULAR GENETICS
Methodology
PCR/Sanger Sequencing
This test utilizes a two-step molecular methodology:
* PCR (Polymerase Chain Reaction):
* Principle: PCR is used to amplify specific regions of the TNF-alpha (TNF-\\alpha) gene that contain the common single nucleotide polymorphisms (SNPs) of interest (e.g., -308G>A, -238G>A, -857C>T, -1031T>C). Primers are designed to flank these polymorphic sites.
* Purpose: To generate millions of copies of the target DNA sequences from the patient's sample, making them sufficient for subsequent sequencing.
* Sanger Sequencing:
* Principle: The amplified PCR products are then subjected to Sanger sequencing. This "chain termination" method uses dideoxynucleotides (ddNTPs) to produce DNA fragments of varying lengths, each terminated at a specific base. These fragments are then separated by size (e.g., via capillary electrophoresis), and a detector reads the fluorescent labels on the ddNTPs, generating a chromatogram (DNA sequence).
* Purpose: To precisely determine the nucleotide sequence at the polymorphic sites within the TNF-alpha gene. This allows for the accurate identification of the specific alleles (e.g., G or A at -308, C or T at -857) present in the patient's DNA.
This combined approach provides high accuracy and specificity for identifying the various single nucleotide polymorphisms (SNPs) within the TNF-alpha gene.
Sample Required
Whole Blood (WB) in EDTA
* Typically, 2-5 mL of Whole Blood collected in an EDTA (lavender-top) vacutainer tube is required. EDTA is an anticoagulant that prevents blood clotting and optimally preserves the DNA within white blood cells.
* Proper collection and timely transport (preferably refrigerated) are crucial to ensure the integrity and quality of the DNA sample for accurate analysis.
Preparation
Proper collection and handling of the blood sample are essential for accurate and reliable genetic testing. * No specific patient preparation is required (e.g., fasting or dietary restrictions). * Sam... Read more
Schedule Report
7 Days
Emergency Report
Yes
Frequently Asked Questions
No data available
Test Description
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