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
The simplest method for generating random-ended dsDNA fragments.
Fragment size can be controlled by adjusting the enzyme concentration.
Fragments generated using dsDNA Shearase Plus are ideal for library construction, Next-Gen sequencing, methylated DNA immunopreciptation (MeDIP, MeDIP-Seq), etc.
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
The simplest method for generating random-ended dsDNA fragments.
Fragment size can be controlled by adjusting the enzyme concentration.
Fragments generated using dsDNA Shearase Plus are ideal for library construction, Next-Gen sequencing, methylated DNA immunopreciptation (MeDIP, MeDIP-Seq), etc.
Product Description
dsDNA Shearase Plus is an endonuclease that cleaves phosphodiester bonds in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini. It has a particularly strong preference for dsDNA and generates random-ended DNA fragments of the desired size in a single step. This enzyme is compatible with low volume inputs, thus minimizing sample loss.
Technical Specifications
Concentration
1 U/µl
Enzyme Inactivation
Heat inactivate enzyme at 65°C for 5 min.
Storage
Store at -20°C for up to 12 months. Avoid repeated freeze/thawing of reagents. Prolonged storage is at ≤ -70°C.
Unit Definition
One unit (1 U) is defined at the amount of enzyme required to convert 250 ng human DNA into DNA fragments in the range of 100-500 bp in 20 minutes at 42°C in total reaction volume in 10 µl.
Yes, we observe that there is more “T” in the second base. Only the first and second base are a little imbalanced, but the nucleotide distribution is balanced from the 3rd base onward.
The genomic DNA: enzyme ratio is the most critical step for dsDNA Shearase Plus reaction. It is necessary to scale up or down proportionally the amount of enzyme when adjusting the DNA input.
5X Reaction Buffer
Volume
2 µl
4 µl
8 µl
Template DNA
Volume
250 ng (x µl)
500 ng (x µl)
1 µg (x µl)
Water
Volume
7 µl – x µl
14 µl – x µl
28 µl – x µl
dsDNA Shearase Plus
Volume
1 µl
2 µl
4 µl
Total Volume
10 µl
20 µl
40 µl
1 U is defined as the amount of enzyme required to convert 250 ng human DNA to 100-500 bp in 20 min (1 U is 2 µl).
dsDNA Shearase Plus is dsDNA specific, thus, RNA will remain intact if there is RNA contamination in the samples. Hybridization between DNA and RNA might occur in RNA contaminated samples, reducing the performance and efficiency of the fragmentation. We recommend using RNA-free DNA as a substrate.
Here are some possible reasons: • The dsDNA Shearase Plus is specific for dsDNA and does not work efficiently for ssDNA. • The performance of the enzyme is significantly affected by RNA contamination present in the samples Check the quality of your DNA sample as DNA/RNA hybridization might occur during the 42°C incubation. • Make sure that enzyme is scaled up or down properly in the reaction.
