NDLI: Enhancement of PCR Amplification of Moderate GC-Containing and Highly GC-Rich DNA Sequences

Content Provider	Springer Nature Link
Author	Strien, Juliane Sanft, Juliane Mall, Gita
Copyright Year	2013
Abstract	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 (MgCl$_{2}$), 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.
Starting Page	1048
Ending Page	1054
Page Count	7
File Format	PDF
ISSN	10736085
Journal	Molecular Biotechnology
Volume Number	54
Issue Number	3
e-ISSN	15590305
Language	English
Publisher	Humana Press Inc
Publisher Date	2013-04-09
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	PCR GC-rich DNA Additive Enhancer DMSO Biotechnology Biochemistry Cell Biology Protein Science Biological Techniques Human Genetics
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biochemistry Bioengineering Applied Microbiology and Biotechnology Biotechnology

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