Difference between revisions of "Boom et al. 1990"
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| + | =Citation= | ||
Boom, R. C. J. A., Sol, C. J., Salimans, M. M., Jansen, C. L., Wertheim-van Dillen, P. M., & Van der Noordaa, J. P. M. E. (1990). Rapid and simple method for purification of nucleic acids. ''[[Journal of Clinical Microbiology]]'', 28(3), 495-503. | Boom, R. C. J. A., Sol, C. J., Salimans, M. M., Jansen, C. L., Wertheim-van Dillen, P. M., & Van der Noordaa, J. P. M. E. (1990). Rapid and simple method for purification of nucleic acids. ''[[Journal of Clinical Microbiology]]'', 28(3), 495-503. | ||
| − | https://www.ncbi.nlm.nih.gov/pubmed/1691208 | + | =Links= |
| − | + | * https://www.ncbi.nlm.nih.gov/pubmed/1691208 | |
| − | http://jcm.asm.org/content/28/3/495.long | + | * http://jcm.asm.org/content/28/3/495.long |
| − | + | * https://scholar.google.com/scholar?cluster=11897106297471650694 | |
| − | https://scholar.google.com/scholar?cluster=11897106297471650694 | + | * * http://hawaiireedlab.com/pdf/boometal1990.pdf (internal lab link only) |
=Published Abstract= | =Published Abstract= | ||
| − | |||
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. | 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. | ||
Revision as of 06:36, 8 September 2018
Citation
Boom, R. C. J. A., Sol, C. J., Salimans, M. M., Jansen, C. L., Wertheim-van Dillen, P. M., & Van der Noordaa, J. P. M. E. (1990). Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology, 28(3), 495-503.
Links
- https://www.ncbi.nlm.nih.gov/pubmed/1691208
- http://jcm.asm.org/content/28/3/495.long
- https://scholar.google.com/scholar?cluster=11897106297471650694
- * http://hawaiireedlab.com/pdf/boometal1990.pdf (internal lab link only)
Published Abstract
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.
