Rapid and simple method for purification of nucleic acids
Bibliographic Data
| ID | 23330355 |
|---|---|
| Authors | René Boom (Amsterdam UMC Location University of Amsterdam), C J Sol (Amsterdam UMC Location University of Amsterdam), C J A Sol (Amsterdam UMC Location University of Amsterdam), M M Salimans (Amsterdam UMC Location University of Amsterdam), C L Jansen (0000-0002-4190-0916, Amsterdam UMC Location University of Amsterdam), Pauline M Wertheim-van Dillen (Amsterdam UMC Location University of Amsterdam), J van der Noordaa (Amsterdam UMC Location University of Amsterdam) |
| Year | 1990 |
| Volume | 28 |
| Issue | 3 |
| Pages | 495-503 |
| Publication date | 1990-03-01 |
| Peer Reviewed | Yes |
| Open Access | Yes |
| Type | ARTICLE |
| Venue | Journal of Clinical Microbiology (JOURNAL) |
| Journal identifiers | ISSN: 0095-1137 • E-ISSN: 1098-660X |
| Publisher | American Society for Microbiology (PUBLISHER • US) |
| DOI | 10.1128/jcm.28.3.495-503.1990 |
| PMID | 1691208 |
| OpenAlex | W2166229810 |
| Language | EN |
| Citations received | 70 |
We have developed a simple, rapid, and reliable protocol for the small-scale purification of DNA and RNA from, e.g., human serum and urine. The method is based on the lysing and nuclease-inactivating properties of the chaotropic agent guanidinium thiocyanate together with the nucleic acid-binding properties of silica particles or diatoms in the presence of this agent. By using size-fractionated silica particles, nucleic acids (covalently closed circular, relaxed circular, and linear double-stranded DNA; single-stranded DNA; and rRNA) could be purified from 12 different specimens in less than 1 h and were recovered in the initial reaction vessel. Purified DNA (although significantly sheared) was a good substrate for restriction endonucleases and DNA ligase and was recovered with high yields (usually over 50%) from the picogram to the microgram level. Copurified rRNA was recovered almost undegraded. Substituting size-fractionated silica particles for diatoms (the fossilized cell walls of unicellular algae) allowed for the purification of microgram amounts of genomic DNA, plasmid DNA, and rRNA from cell-rich sources, as exemplified for pathogenic gram-negative bacteria. In this paper, we show representative experiments illustrating some characteristics of the procedure which may have wide application in clinical microbiology.
Biology · Chaotropic agent · Chromatography · DNA · DNA ligase · Gene · Ligase chain reaction · Lysis · Nuclease · Nucleic acid · Plasmid · Polymerase chain reaction · Proteinase K · Ribosomal RNA · RNA · Bacteriophages and microbial interactions · Biochemistry · Chemistry · Enterobacteriaceae and Cronobacter Research · Genomics and Phylogenetic Studies · Molecular Biology
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| Unique citing works | 70 |
|---|---|
| Citations per year | 2,12 |
| Citation span | 1993 - 2026 (34) |
| Citation velocity | current |
| Highly cited | No |
| Citation types | Neutral: 57 |