ArticleMolecular biotechnology2013
Amplification of GC-rich DNA for high-throughput family-based genetic studies.
Article in Molecular biotechnology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Optimizing PCR amplification of GC-rich nicotinic acetylcholine receptor subunits from invertebrates.Biochemistry and biophysics reports · 2025Article
- Genetic andFrontiers in genetics · 2025Article
- Engineering a synthesis-friendly serum responsive promoter for antibiotic selection of genomic integration in cell-based therapy.Biotechnology letters · 2022Article
- Design of a synthesis-friendly hypoxia-responsive promoter for cell-based therapeutics.Engineering in life sciences · 2021Article
- Creation of a synthesis-friendly inflammation-inducible promoter suitable for cell therapy.Integrative biology : quantitative biosciences from nano to macro · 2021Article
- Single-nucleotide polymorphism detecting of some candidate genes related to lipid metabolism in Booroola Merino-Afshari sheep by Bayesian model averaging.Tropical animal health and production · 2021Article
- Next-generation sequencing for diagnosis of thoracic aortic aneurysms and dissections: diagnostic yield, novel mutations and genotype phenotype correlations.Journal of translational medicine · 2016Article
- DFT investigation of the vibrational properties of GC Watson-Crick and Hoogsteen base pairs in the presence of Mg²⁺, Ca²⁺, and Cu²⁺ ions.Journal of molecular modeling · 2014Article
- The c.42_52del11 mutation in TPRN and progressive hearing loss in a family from Pakistan.Biochemical genetics · 2013Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors.
Funding
Abstract
Researchers face a significant problem in PCR amplification of DNA fragments with high GC contents. Analysis of these regions is of importance since many regulatory regions of different genes and their first exons are GC-rich. There are a large number of protocols for amplification of GC-rich DNA, some of which perform well but are costly. Most of the economical protocols fail to perform consistently, especially on products with >80 % GC contents and a size of >300 bp. One of these protocols requires multiple additions of DNA polymerase during thermal cycling which therefore rules out its utility if a large number of samples have to be amplified. We have established a method for simultaneous amplification of specific PCR products from a large number of human DNA samples using general laboratory reagents. These amplicons have GC contents ranging from 65-85 % and sizes up to 870 bp. The protocol uses a PCR buffer containing co-solvents including 2-mercaptoethanol and bovine serum albumin for amplification of DNA. A specific thermal cycling profile is also used which incorporates a high annealing temperature in the first 7 cycles of the reactions. The PCR products are suitable for different molecular biology applications including sequencing.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.