Zymo Research는 1994년도에 설립된 미국 회사로 고품질 키트 제조회사 입니다.
각종 샘플(세포, 조직, 환경샘플 등)로부터 고품질
DNA나 RNA를 가장 쉽고 빠르게 뽑을 수 있습니다.
이 외에도, 각종 Epigenetics 관련 제품들
(DNA Methylation kit 등)과 Microbiomics
(샘플 채집부터 분석까지의 전 단계의 제품)
연관 제품들이 준비되어 있습니다.
Highlights
For the isolation of nucleosome-associated DNA from mammalian and yeast cells.
Ideal for use in nucleosome mapping studies.
서한형 대리
Zymo Research 제품 담당자
경신과학(주)
영업부
H.P) 010-8832-6303
HanHyung Seo
Zymo Research Brand Manager
Kyongshin scientific Co., Ltd.
Sales Department
H.P) 82)10-8832-6303
제품소개
Highlights
For the isolation of nucleosome-associated DNA from mammalian and yeast cells.
Ideal for use in nucleosome mapping studies.
Product Description
The EZ Nucleosomal DNA Prep Kit is a streamlined procedure for the isolation of nucleosome-associated DNA. The kit includes reagents/procedures for the following: cell nuclei isolation, intact nuclei enzymatic digestion, and nucleosomal DNA purification. This kit includes two different enzymes for nucleosomal DNA preparation: Atlantis dsDNase and Micrococcal Nuclease. Enzymatic digestion yields very homogeneous populations of core nucleosomes and purification of the nucleosome-associated DNA is performed using Zymo Research’s proven spin column technology. The result is pure nucleosomal DNA ready for analysis in less than 45 minutes!
Technical Specifications
Applicable For
All sensitive downstream applications such as qPCR and Next-Generation sequencing.
Processing Volume
≤106 cells
Purity
Typical A260/A280 ≥ 1.8
Sample Source
Nucleosome associated DNA isolation and purification from mammalian and yeast cells.
Sample Storage
Eluted DNA should be stored at ≤ -20°C.
Yield
Up to 25 µg total DNA can be eluted into ≥ 25 µl. For DNA 75 bp - 10 kb the recovery is 70-90%. For DNA 11 kb - 23 kb the recovery is 50-70%.
Once the nuclei are isolated, the enzyme sensitivity should be similar between different cell types. However, some cell lines will be more sensitive to the detergent in the Nuclei Prep Buffer and thus result in a loss of nuclei. You can dilute the Nuclei Prep Buffer 1:1 with dsDNase Digestion Buffer prior to nuclei isolation.
• Atlantis dsDNase is a double-stranded DNA specific endonuclease that cleaves phosphodiester bonds in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini. Increasing the incubation time with Atlantis dsDNAse will increase efficiency and generate more mono-nucleosomal DNA. • Micrococcal Nuclease (MNase) is a single and double-stranded DNA and RNA endonuclease. MNase is more specific for single-stranded nucleic acids, but cleavage is biased towards AT-rich and AU-rich sequences. MNase will result in more robust digestion compared to Atlantis dsDNase. Increasing the incubation time with MNase will increase efficiency of this enzyme and generate more mono-nucleosomal DNA. • The enzyme selection depends on desired downstream applications.
Washing the trypsinized cell pellets with PBS is very critical step. Residual EDTA in the cell pellets will decrease both Atlantis dsDNAse and MNase digestion efficiency. Other considerations for incomplete shearing: • Check that the correct number of cells was digested. (D5220 kit: 1x106 mammalian cells) • If you only want mono-nucleosomal DNA instead of nucleosomal ladder containing mono-, di-, tri-nucleosomal, etc., you will need to increase the enzyme concentration. Based on our experience, we need to use >1U per Atlantis dsDNAse per million cells.
