dsDNA Shearase Plus
E2018-50 / E2018-200
- 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.
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.
|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.|
Q1: Is there any sequence preference at the cutting sites?
Q2: Can reaction be scaled up/down?
Q3: How can I ensure the enzyme has been inactivated?
Q4: Can AMPure beads be used to clean up the reaction?
Q5: Can RNA be used as a substrate for dsDNA Shearase Plus?
Q6: Is the enzymatic digestion affected by DNA methylation?
Q7: Why is the enzyme not shearing properly?
“I believe our whole lab wants to switch to your shearase to put an end to all the sonicating we are currently doing for ChIP, MeDIP, and MeDIP-seq experiments.”
- Garrett K (The University of Alabama at Birmingham)
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