MilliporeSigma
  • Home
  • Search Results
  • Identification of RNase-resistant RNAs in Saccharomyces cerevisiae extracts: Separation from chromosomal DNA by selective precipitation.

Identification of RNase-resistant RNAs in Saccharomyces cerevisiae extracts: Separation from chromosomal DNA by selective precipitation.

Analytical biochemistry (2015-09-30)
Blanca V Rodriguez, Eric T Malczewskyj, Joshua M Cabiya, L Kevin Lewis, Corina Maeder
ABSTRACT

High-quality chromosomal DNA is a requirement for many biochemical and molecular biological techniques. To isolate cellular DNA, standard protocols typically lyse cells and separate nucleic acids from other biological molecules using a combination of chemical and physical methods. After a standard chemical-based protocol to isolate chromosomal DNA from Saccharomyces cerevisiae and then treatment with RNase A to degrade RNA, two RNase-resistant bands persisted when analyzed using gel electrophoresis. Interestingly, such resistant bands did not appear in preparations of Escherichia coli bacterial DNA after RNase treatment. Several enzymatic, chemical, and physical methods were employed in an effort to remove the resistant RNAs, including use of multiple RNases and alcohol precipitation, base hydrolysis, and chromatographic methods. These experiments resulted in the development of a new method for isolation of S. cerevisiae chromosomal DNA. This method utilizes selective precipitation of DNA in the presence of a potassium acetate/isopropanol mixture and produces high yields of chromosomal DNA without detectable contaminating RNAs.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Potassium acetate, BioXtra, ≥99.0%
Sigma-Aldrich
Potassium acetate, meets USP testing specifications
Sigma-Aldrich
Potassium acetate, for molecular biology, ≥99.0%
Sigma-Aldrich
Sodium hydroxide-16O solution, 20 wt. % in H216O, 99.9 atom % 16O
Sigma-Aldrich
Boric acid, ACS reagent, ≥99.5%
Sigma-Aldrich
Boric acid, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Potassium acetate, puriss., meets analytical specification of Ph. Eur., BP, E261, 99-101%
Sigma-Aldrich
Boric acid, puriss., meets analytical specification of Ph. Eur., BP, NF, 99.5-100.5%, powder
Sigma-Aldrich
Potassium acetate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.0%
Sigma-Aldrich
Potassium acetate, ACS reagent, ≥99.0%
Sigma-Aldrich
Potassium acetate, ReagentPlus®, ≥99.0%
SAFC
Tromethamine
Sigma-Aldrich
Boric acid, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., buffer substance, ≥99.8%
Sigma-Aldrich
Boric acid, tablet, 1 g boric acid per tablet
Sigma-Aldrich
Boric acid, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
Boric acid, BioXtra, ≥99.5%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, 99.995% trace metals basis
Sigma-Aldrich
Boric acid, 99.999% trace metals basis
Sigma-Aldrich
Potassium acetate, 98%
Sigma-Aldrich
Tromethamine, meets USP testing specifications
Sigma-Aldrich
Sigma 7-9®, ≥99% (titration), crystalline
Sigma-Aldrich
Boric acid, suitable for electrophoresis, ≥99.5%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)
Sigma-Aldrich
Boric acid, 99.97% trace metals basis
Sigma-Aldrich
Boric acid, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
Potassium acetate, BioUltra, for molecular biology, ≥99.0% (NT)
Sigma-Aldrich
Potassium acetate, 99.98% trace metals basis