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
Ideal for use in nucleosome mapping studies.
Sold separately or as part of EZ Nucleosomal DNA Prep Kit (D5220).
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
Ideal for use in nucleosome mapping studies.
Sold separately or as part of EZ Nucleosomal DNA Prep Kit (D5220).
Product Description
Atlantis dsDNase is a double-stranded DNA-specific endonuclease that cleaves phosphodiester bonds in DNA to yield homogeneous populations of core nucleosomes.
Technical Specifications
Concentration
0.1 U/µl
Inactivation
5X MN Stop Buffer or EDTA
Standard Reaction Time
20 minutes
Storage
Store at -20°C for up to 12 months. Avoid repeated freeze/thawing. Prolonged storage should be ≤-70°C.
Unit Definition
One unit (U) is defined as the amount of enzyme needed to produce an increase in absorbance at 260 nm of 0.001 per minute, using 50 mg/ml high MW DNA in 50 mM Na-acetate pH 5.0 and 5 mM MgCl2 (Kunitz, 1950).
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.
