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Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity

Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity

Optimizing the polymerase chain reaction (PCR) continues to be a major challenge in molecular biology, and obtaining the correct magnesium chloride (MgCl2) concentration is key to a successful reaction. A clear understanding of how MgCl2 affects PCR thermodynamics and kinetics is crucial for creatin...

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में बचाया:
ग्रंथसूची विवरण
मुख्य लेखकों: Povetkin, S. N., Поветкин, С. Н.
स्वरूप: Статья
भाषा:English
प्रकाशित: Academic Press Inc. 2025
विषय:
DNA melting temperature
Thermodynamics of PCR
PCR optimization
PCR efficiency
Magnesium chloride (MgCl<sub>2</sub>)
ऑनलाइन पहुंच:https://dspace.ncfu.ru/handle/123456789/30526
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spelling ir-123456789-305262025-06-19T08:24:19Z Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity Povetkin, S. N. Поветкин, С. Н. DNA melting temperature Thermodynamics of PCR PCR optimization PCR efficiency Magnesium chloride (MgCl<sub>2</sub>) Optimizing the polymerase chain reaction (PCR) continues to be a major challenge in molecular biology, and obtaining the correct magnesium chloride (MgCl2) concentration is key to a successful reaction. A clear understanding of how MgCl2 affects PCR thermodynamics and kinetics is crucial for creating efficient and reliable protocols that work consistently. A systematic meta-analysis was conducted of 61 peer-reviewed studies published between 1973 and 2024. The study selection adhered to rigorous PICOS criteria, prioritizing experimental investigations that specifically examined the effects of magnesium chloride (MgCl2) on key PCR parameters. Data extraction and subsequent analyses were performed using standardized methodologies, with particular emphasis on template characteristics, reaction conditions, and their interplay in influencing PCR efficiency and specificity. The analysis showed a strong logarithmic relationship between MgCl2 concentration and DNA melting temperature, with an optimal ranges of 1.5 and 3.0 mM. Every 0.5 mM increase in MgCl2 within this range was associated with a 1.2 °C increase in melting temperature. Template complexity significantly affected the optimal MgCl2 requirements, with genomic DNA templates requiring higher concentrations than the more straightforward templates. This meta-analysis offers quantitative insights and evidence-based guidelines for optimizing magnesium chloride (MgCl2) concentration using polymerase chain reaction (PCR). These results demonstrate that the precise modulation of MgCl2 concentration, tailored to specific template characteristics, can significantly improve both the efficiency and specificity of PCR. These findings provide a robust theoretical framework for the development of template-specific optimization strategies and advance the design of more reliable and effective PCR protocols. 2025-06-19T08:23:18Z 2025-06-19T08:23:18Z 2025 Статья Tbahriti H.F., Zerrouki A., Boukadoum A., Kameche M., Povetkin S., Simonov A., Thiruvengadam M. Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity // Analytical Biochemistry. - 2025. - 705. - art. no. 115909. - DOI: 10.1016/j.ab.2025.115909 https://dspace.ncfu.ru/handle/123456789/30526 en Analytical Biochemistry application/pdf application/pdf Academic Press Inc.
institution СКФУ
collection Репозиторий
language English
topic DNA melting temperature
Thermodynamics of PCR
PCR optimization
PCR efficiency
Magnesium chloride (MgCl<sub>2</sub>)
spellingShingle DNA melting temperature
Thermodynamics of PCR
PCR optimization
PCR efficiency
Magnesium chloride (MgCl<sub>2</sub>)
Povetkin, S. N.
Поветкин, С. Н.
Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity
description Optimizing the polymerase chain reaction (PCR) continues to be a major challenge in molecular biology, and obtaining the correct magnesium chloride (MgCl2) concentration is key to a successful reaction. A clear understanding of how MgCl2 affects PCR thermodynamics and kinetics is crucial for creating efficient and reliable protocols that work consistently. A systematic meta-analysis was conducted of 61 peer-reviewed studies published between 1973 and 2024. The study selection adhered to rigorous PICOS criteria, prioritizing experimental investigations that specifically examined the effects of magnesium chloride (MgCl2) on key PCR parameters. Data extraction and subsequent analyses were performed using standardized methodologies, with particular emphasis on template characteristics, reaction conditions, and their interplay in influencing PCR efficiency and specificity. The analysis showed a strong logarithmic relationship between MgCl2 concentration and DNA melting temperature, with an optimal ranges of 1.5 and 3.0 mM. Every 0.5 mM increase in MgCl2 within this range was associated with a 1.2 °C increase in melting temperature. Template complexity significantly affected the optimal MgCl2 requirements, with genomic DNA templates requiring higher concentrations than the more straightforward templates. This meta-analysis offers quantitative insights and evidence-based guidelines for optimizing magnesium chloride (MgCl2) concentration using polymerase chain reaction (PCR). These results demonstrate that the precise modulation of MgCl2 concentration, tailored to specific template characteristics, can significantly improve both the efficiency and specificity of PCR. These findings provide a robust theoretical framework for the development of template-specific optimization strategies and advance the design of more reliable and effective PCR protocols.
format Статья
author Povetkin, S. N.
Поветкин, С. Н.
author_facet Povetkin, S. N.
Поветкин, С. Н.
author_sort Povetkin, S. N.
title Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity
title_short Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity
title_full Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity
title_fullStr Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity
title_full_unstemmed Comprehensive review and meta-analysis of magnesium chloride optimization in PCR: Investigating concentration effects on reaction efficiency and template specificity
title_sort comprehensive review and meta-analysis of magnesium chloride optimization in pcr: investigating concentration effects on reaction efficiency and template specificity
publisher Academic Press Inc.
publishDate 2025
url https://dspace.ncfu.ru/handle/123456789/30526
work_keys_str_mv AT povetkinsn comprehensivereviewandmetaanalysisofmagnesiumchlorideoptimizationinpcrinvestigatingconcentrationeffectsonreactionefficiencyandtemplatespecificity
AT povetkinsn comprehensivereviewandmetaanalysisofmagnesiumchlorideoptimizationinpcrinvestigatingconcentrationeffectsonreactionefficiencyandtemplatespecificity
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