Journal
EXPERT REVIEW OF MOLECULAR DIAGNOSTICS
Volume 5, Issue 5, Pages 809-815Publisher
TAYLOR & FRANCIS AS
DOI: 10.1586/14737159.5.5.809
Keywords
aging; allelic; preference; jumping PCR; mitochondrial DNA; mutational spectra; PCR errors; single-molecule PCR; somatic mutations; recombination
Categories
Funding
- NIA NIH HHS [AG19787] Funding Source: Medline
- NIEHS NIH HHS [ES11343] Funding Source: Medline
- NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES [R01ES011343] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE ON AGING [R01AG019787] Funding Source: NIH RePORTER
Ask authors/readers for more resources
A critical review of the clone-by-clone approach to the analysis of complex spectra of somatic mutations is presented. The study of a priori unknown somatic mutations requires painstaking analysis of complex mixtures of multiple mutant and non-mutant DNA molecules. If mutant fractions are sufficiently high, these mixtures can be dissected by the cloning of individual DNA molecules and scanning of the individual clones for mutations (e.g., by sequencing). Currently, the majority of such cloning is performed using PCR fragments. However, post-PCR cloning may result in various PCR artifacts - PCR errors and jumping PCR - and preferential amplification of certain mutations. This review argues that single-molecule PCR is a simple alternative that promises to evade the disadvantages inherent to post-PCR cloning and enhance mutational analysis in the future.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available