Methylation-specific PCR: four steps in primer design
Апстракт
Methylation-specific PCR (MSP) is still the method of choice for a single gene methylation study. The proper design of the primer pairs is a prerequisite for obtaining reliable PCR results. Despite numerous protocols describing the rules for MSP primer design, none of them provide a comprehensive approach to the problem. Our aim was to depict a workflow for the primer design that is concise and easy to follow. In order to achieve this goal, adequate tools for promoter sequence retrieval, MSP primer design and subsequent in silico analysis are presented and discussed. Furthermore, a few instructive examples regarding a good versus a poor primer design are provided. Finally, primer design is demonstrated according to the proposed workflow. This article aims to provide researchers, interested in a single gene methylation studies, with useful information regarding successful primer design.
Кључне речи:
Methylation-specific PCR / Primer Design / Database / Software for MSP Primer Design / Transcriptional Start SiteИзвор:
Central European Journal of Biology, 2014, 9, 12, 1127-1139Финансирање / пројекти:
- Молекуларне детерминанте за дизајн тумор маркера (RS-MESTD-Basic Research (BR or ON)-173049)
DOI: 10.2478/s11535-014-0324-z
ISSN: 1895-104X; 1644-3632
WoS: 000340681200001
Scopus: 2-s2.0-84906303114
Колекције
Институција/група
VinčaTY - JOUR AU - Davidović, Radoslav S. AU - Božović, Ana M. AU - Mandušić, Vesna AU - Krajnović, Milena M. PY - 2014 UR - https://vinar.vin.bg.ac.rs/handle/123456789/74 AB - Methylation-specific PCR (MSP) is still the method of choice for a single gene methylation study. The proper design of the primer pairs is a prerequisite for obtaining reliable PCR results. Despite numerous protocols describing the rules for MSP primer design, none of them provide a comprehensive approach to the problem. Our aim was to depict a workflow for the primer design that is concise and easy to follow. In order to achieve this goal, adequate tools for promoter sequence retrieval, MSP primer design and subsequent in silico analysis are presented and discussed. Furthermore, a few instructive examples regarding a good versus a poor primer design are provided. Finally, primer design is demonstrated according to the proposed workflow. This article aims to provide researchers, interested in a single gene methylation studies, with useful information regarding successful primer design. T2 - Central European Journal of Biology T1 - Methylation-specific PCR: four steps in primer design VL - 9 IS - 12 SP - 1127 EP - 1139 DO - 10.2478/s11535-014-0324-z ER -
@article{ author = "Davidović, Radoslav S. and Božović, Ana M. and Mandušić, Vesna and Krajnović, Milena M.", year = "2014", abstract = "Methylation-specific PCR (MSP) is still the method of choice for a single gene methylation study. The proper design of the primer pairs is a prerequisite for obtaining reliable PCR results. Despite numerous protocols describing the rules for MSP primer design, none of them provide a comprehensive approach to the problem. Our aim was to depict a workflow for the primer design that is concise and easy to follow. In order to achieve this goal, adequate tools for promoter sequence retrieval, MSP primer design and subsequent in silico analysis are presented and discussed. Furthermore, a few instructive examples regarding a good versus a poor primer design are provided. Finally, primer design is demonstrated according to the proposed workflow. This article aims to provide researchers, interested in a single gene methylation studies, with useful information regarding successful primer design.", journal = "Central European Journal of Biology", title = "Methylation-specific PCR: four steps in primer design", volume = "9", number = "12", pages = "1127-1139", doi = "10.2478/s11535-014-0324-z" }
Davidović, R. S., Božović, A. M., Mandušić, V.,& Krajnović, M. M.. (2014). Methylation-specific PCR: four steps in primer design. in Central European Journal of Biology, 9(12), 1127-1139. https://doi.org/10.2478/s11535-014-0324-z
Davidović RS, Božović AM, Mandušić V, Krajnović MM. Methylation-specific PCR: four steps in primer design. in Central European Journal of Biology. 2014;9(12):1127-1139. doi:10.2478/s11535-014-0324-z .
Davidović, Radoslav S., Božović, Ana M., Mandušić, Vesna, Krajnović, Milena M., "Methylation-specific PCR: four steps in primer design" in Central European Journal of Biology, 9, no. 12 (2014):1127-1139, https://doi.org/10.2478/s11535-014-0324-z . .