ArticleJournal of clinical laboratory analysis2013
Optimization of PCR conditions for amplification of GC-Rich EGFR promoter sequence.
Article in Journal of clinical laboratory analysis, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 49 citations in OpenAlex.
- Optimization of PCR amplification and sequencing of GC-rich protein-coding region of CEBPA (CCAAT/enhancer binding protein alpha) in acute myeloid leukemia patients using betaine.Molecular biology reports · 2026Article
- Genomic Variant Surveillance of SARS-CoV-2 Positive Specimens Using a Direct PCR Product Sequencing Surveillance (DPPSS) Method.Acta medica Philippina · 2026Article
- Comparative Analysis of Two Digital PCR Platforms for Detecting DNA Methylation in Patient Samples.Cell biochemistry and function · 2025Article
- Article
- Investigating the frequencies of EGFR mutations and EGFR single nucleotide polymorphisms genotypes and their predictive role in NSCLC patients in Republic of Serbia.Molecular biology reports · 2025Article
- Modulation of mechanosensitive genes during embryonic aortic arch development.Developmental dynamics : an official publication of the American Association of Anatomists · 2025Article
- Zoonotic filariasis and its public health significance: a comprehensive literature review.Frontiers in microbiology · 2025Review
- Optimization of 18 S rRNA metabarcoding for the simultaneous diagnosis of intestinal parasites.Scientific reports · 2024Article
- Efficient cloning of linear DNA inserts (ECOLI) into plasmids using site-directed mutagenesis.Scientific reports · 2024Article
- Implementing L-DNA analogs as mirrors of PCR reactant hybridization state: theoretical and practical guidelines for PCR cycle control.Analytical methods : advancing methods and applications · 2024Article
- PCR in Forensic Science: A Critical Review.Genes · 2024Review
- Evolution of dirofilariasis diagnostic techniques from traditional morphological analysis to molecular-based techniques: a comprehensive review.Frontiers in parasitology · 2024Review
- Significance of EGFR investigation in odontogenic keratocyst: a narrative review.Molecular biology reports · 2023Review
- Response Surface Methodology for Optimization of Multiplex-PCR Protocols for Detection of TYLCV, TSWV andGenes · 2023Article
- Using selenocysteine-specific reporters to screen for efficient tRNAMethods in enzymology · 2022Article
- Primer Designing for Amplifying an AT-Rich Promoter from Arabidopsis thaliana.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Development and application of multiplex PCR method for simultaneous detection of seven viruses in ducks.BMC veterinary research · 2019Article
- Epidermal Growth Factor Receptor Gene in Non-Small-Cell Lung Cancer: The Importance of Promoter Polymorphism Investigation.Analytical cellular pathology (Amsterdam) · 2018Review
- Optimization of PCR conditions for amplifying an AT-rich amino acid transporter promoter sequence with high number of tandem repeats from Arabidopsis thaliana.BMC research notes · 2017Article
- Frequencies of EGFR single nucleotide polymorphisms in non-small cell lung cancer patients and healthy individuals in the Republic of Serbia: a preliminary study.Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine · 2016Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundPolymerase chain reaction (PCR) is an extremely sensitive method that often demands optimization, especially when difficult templates need to be amplified. The aim of the present study was to optimize the PCR conditions for amplification of the epidermal growth factor receptor (EGFR) promoter sequence featuring an extremely high guanine-cytosine (GC) content in order to detect single nucleotide polymorphisms -216G>T and -191C>A.
methodsGenomic DNA used for amplification was extracted from formalin-fixed paraffin-embedded lung tumor tissue and PCR products were detected by agarose gel electrophoresis.
resultsResults showed that addition of 5% dimethyl sulfoxide (DMSO), as well as DNA concentration in PCR reaction of at least 2 μg/ml, were necessary for successful amplification. Due to high GC content, optimal annealing temperature was 7°C higher than calculated, while adequate MgCl2 concentration ranged from 1.5 to 2.0 mM.
conclusionIn conclusion, EGFR promoter region is a difficult PCR target, but it could be amplified after optimization of MgCl2 concentration and annealing temperature in the presence of DMSO and the DNA template of acceptable concentration.
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