Enhancement of PCR Amplification of Moderate GC-Containing and Highly GC-Rich DNA Sequences
| Autor: | Gita Mall, Juliane Sanft, Juliane Strien |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
GC Rich Sequence
Magnesium Chloride Bioengineering Polymerase Chain Reaction Applied Microbiology and Biotechnology Biochemistry law.invention chemistry.chemical_compound law Primer dimer Humans Dimethyl Sulfoxide Bovine serum albumin Molecular Biology Polymerase Polymerase chain reaction DNA Primers Base Composition Formamides biology Multiple displacement amplification Serum Albumin Bovine DNA chemistry biology.protein Primer (molecular biology) Biotechnology |
| Zdroj: | Molecular Biotechnology. 54:1048-1054 |
| ISSN: | 1559-0305 1073-6085 |
| DOI: | 10.1007/s12033-013-9660-x |
| Popis: | PCR is a commonly used and highly efficient technique in biomolecular laboratories for specific amplification of DNA. However, successful DNA amplification can be very time consuming and troublesome because many factors influence PCR efficiency. Especially GC-rich DNA complicates amplification because of generation of secondary structures that hinder denaturation and primer annealing. We investigated the impact of previously recommended additives such as dimethylsulfoxide (DMSO), magnesium chloride (MgCl2), bovine serum albumin (BSA), or formamide. Furthermore, we tested company-specific substances as Q-Solution, High GC Enhancer, and Hi-Spec; various actively promoted polymerases as well as different PCR conditions for their positive effects on DNA amplification of templates with moderate and extremely high CG-content. We found considerable differences of specificity and quantity of product between different terms. In this article, we introduce conditions for optimized PCR to help resolve problems amplifying moderate to high GC-rich templates. |
| Databáze: | OpenAIRE |
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