About EZ Nucleosomal DNA Prep Kit
The EZ Nucleosomal DNA Prep Kit is a streamlined procedure for the isolation of mammalian and yeast nucleosome-associated DNA. The kit includes procedures and reagents for: cell nuclei isolation, intact nuclei micrococcal nuclease digestion, and nucleosomal DNA purification. Non-nucleosomal DNA is specifically degraded using micrococcal nuclease and an optimized reaction buffer; while purification of "protected" nucleosome-associated DNA is performed using Zymo Research's proven Fast-Spin column technology. The result is pure nucleosomal DNA ready for analysis in less than 30 minutes!
|DNA Recovery||Typically, up to 25 µg total DNA can be eluted into as little as 25 µl water. For DNA 75 bp to 10 kb the recovery is 70-90%. For DNA 11 kb to 23 kb the recovery is 50-70%.|
|Sample Sources||Nucleosome associated DNA isolation and purification from mammalian and yeast cells.|
|DNA Purity||High quality, purified DNA is eluted with elution buffer or water and is especially well suited for PCR amplification, arrays, Southern blot analysis, DNA quantification, sequencing, and other molecular applications.|
|Product Detergent Tolerance||≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤0.1% SDS|
Mammalian Nucleosomal DNA Preparation: Mammalian nuclei was treated with 0.1 U, 0.25 U, and 0.5 U (unit) Atlantis dsDNase for the 20 min at 42°C. DNA was subsequently resolved in a 2% agarose gel. 100 bp DNA ladder (Zymo Research Corp.). Asterisks (1N, 2N, 3N) represent mono-, di-, and tri-nucleosomal DNAs, respectively.
Mononucleosomal DNA from p16-methylated AGS and p16-unmethylated MGC803 cell lines was isolated using the EZ Nucleosomal DNA Prep Kit. The isolated mononucleosomal DNA was used to study the chromatin accessibility across the p16 promoter by quantitative PCR.