+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Comparison of two methods for the small-scale extraction of DNA from subgingival microorganisms

, : Comparison of two methods for the small-scale extraction of DNA from subgingival microorganisms. Oral Microbiology and Immunology 4(3): 135-140

Two methods were compared for the extraction of DNA from small numbers of bacterial cells. The first method involved lysis of cells with SDS in the presence of proteinase K, treatment with hexadecyltrimethyl ammonium bromide (CTAB) and precipitation of DNA with isopropanol. In the second method, DNA was extracted by treatment of the cells with guanidine hydrochloride (GHCl) and precipitated with ethanol. Thirty strains of representative gram positive and gram negative species were included in the study. Preparations derived from confluent growth on one-quarter of the surface of agar plates and from 10(8) cells were subjected to each extraction procedure and analyzed for their content of DNA, RNA and protein. The suitabilities of the resultant DNA for restriction enzyme digestion and biotin-labelling by a random primer technique were also assessed. In general, the CTAB method yielded greater amounts of DNA than the GHCl procedure. RNA was present in most preparations of both types, but in amounts detectable only by agarose gel electrophoresis. The latter technique also revealed that DNA was not excessively sheared by either procedure. Protein was detected in some CTAB and GHCl preparations, but was not consistently associated with one or the other method. DNA obtained by both methods could be digested by the restriction enzyme EcoR I. In addition, biotin-labelled DNA probes prepared from CTAB and GHCl preparations were capable of hybridizing with homologous target DNA fixed to nitrocellulose. Since the CTAB method was consistently successful in recovering DNA from preparations containing 10(8) cells, it may be more suitable for the direct treatment of single colonies taken from primary isolation plates or plaque samples.


PMID: 2639297

Other references

Misra N., 1989: Studies on fungi deteriorating stored fruits of emblica officinalis gaertn. Thirteen fungi were isolated from stored fruits of Emblica officinalis Gaertn using standard blotter and agar plate methods. Nine fungi appeared by using both the methods. Ten were isolated both from surface unsterilized and surface sterilized fru...

Ghosh, S.; Natarajan, R., 2001: Cloning of the human cholesteryl ester hydrolase promoter: identification of functional peroxisomal proliferator-activated receptor responsive elements. Cholesteryl ester hydrolase (CEH) is responsible for hydrolysis of stored cholesterol esters in macrophage foam cells and release of free cholesterol for high-density lipoprotein-mediated efflux. PCR-based screening of human genomic libraries with...

Walter, S.J.; Unwin, R.J.; Shirley, D.G., 1997: Effect of barium on sodium reabsorption in perfused loops of Henle in anaesthetized rats. Journal of Physiology (Cambridge) 501P(0): 164P-165P

Chey, W.Y.; Chang, T.M.; Lee, K.Y.; Song, Y.; Li, P.J.; Goldstein, J.; Decross, A., 2000: Diagnostic challenge in patients with duodenal ulcerogenic islet cell tumors. Pancreas 21(4): 436, November

Nehring, A., 1901: Ueber Dipus schluetteri, n. sp. und einige andere Nager aus Palastina. Sitzungsberichte der Gesellschaft Naturforschender zu Berlin, 163-176

Chauris L., 1978: Succession of intrusions in the upper Variscan granitic complex of Morlaix Bay, Armorican Massif. Comptes Rendus Hebdomadaires des Seances de l'Academie des Sciences, Serie D: Sciences Naturelles 286(3): 253-256

Tamagawa, H.; Takahashi, I.; Furuse, M.; Yoshitake Kitano, Y.; Tsukita, S.; Ito, T.; Matsuda, H.; Kiyono, H., 2003: Characteristics of claudin expression in follicle-associated epithelium of Peyer's patches: Preferential localization of claudin-4 at the apex of the dome region. Gut-associated lymphoreticular tissues, such as Peyer's patches and cecal patches, are important inductive sites for mucosal immune responses. As such, gut-associated lymphoreticular tissues may have an epithelial barrier different from that...

Stulianea Fountoulake, A.; Bisbike Demetriadou, A.; Kakkinou Aneste, D.; Mpathrellos, N.; Papaemmanoule, B.; Spuropoulou, P.N.; Paulakou, B., 1984: Study of biochemical characters of Aspergillus isolates from pathological material. In studies with A. fumigatus, A. nidulans, A. flavus, A. glaucus, A. jeanselmei, A. niger and A. clavatus, differences were observed in sugar fermentation and the use of carbon and nitrogen sources by the spp.

Bertin, C.Helena.Figueiredo.Preza.; Rezende, M.Andrade.; Sigulem, D.Maria.; Morais, T.Beninga., 2009: Hurdles at work: perceptions of hospital food handlers. Food handlers have a very important role in preventing food contamination during its preparation and distribution. This responsibility is even greater in hospitals, since a large number of patients have low immunity and consequently food contamina...

Wang, J.; Yang, W.; Tang, Y.; Xu, Q.; Huang, S.; Yao, J.; Zhang, J.; Lei, Z., 2016: Regioselective sulfation of Artemisia sphaerocephala polysaccharide: Solution conformation and antioxidant activities in vitro. Regioselective modification is an effective approach to synthesize polysaccharides with different structure features and improved properties. In this study, regioselective sulfation of Artemisia sphaerocephala polysaccharide (SRSASP) was prepared...