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Epigenetics |
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| Epigenetics>【Zymo】 |
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dsDNA Shearase™ Plus（E2018） |
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dsDNA Shearase™ Plus
About dsDNA Shearase™ Plus
dsDNA Shearase™ Plus is an endonuclease that cleaves phosphodiester bonds in DNA to yield oligonucleotides with 5'-phosphates 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.
| Unit Definition |
One unit (U) is defined as 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 a total reaction volume of 10 µl. |
| Specificity |
Provided with 5X dsDNA Shearase™ Plus Reaction Buffer |
| Enzyme Inactivation and DNA Cleanup |
For enzyme inactivation and DNA purification we recommend the DNA Clean & Concentrator™ kit. Addition of 5 volumes of DNA Binding Buffer from the DNA Clean & Concentrator™ kit to each reaction volume (5:1) inactivates dsDNA Shearase™ Plus. Alternatively, the enzyme can be inactivated by incubating at 65°C for 5 minutes followed by DNA purification by phenol/chloroform extraction. |
| Enzyme Concentration |
1 U/µl |
| Enzyme Inactivation |
65°C for 5 min. |
| Optimum Reaction Temperature |
42°C |
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| Fragmentation of HCT116 Cell DNA Using dsDNA Shearase™ Plus. 250 ng or 500 ng of HCT116 cell gDNA was incubated with 1, 0.5, 0.25, or 0.1 U dsDNA Shearase™ Plus for 20 min at 42°C. The reaction was stopped by incubating at 65°C for 5 min. Fragmented DNA was purified with the DNA Clean & Concentrator™ kit and subsequently resolved in a 1% agarose gel. |
The simplest method for generating random-ended dsDNA fragments.
Fragment size is conveniently controlled by adjusting the enzyme concentration.
dsDNA Shearase™ Plus-generated fragments are ideal for library construction, Next-Gen sequencing, and methylated DNA immunoprecipitation (MeDIP).
| dsDNA Shearase™ Plus (50 U) |
50 Units |
E2018-50 |
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| dsDNA Shearase™ Plus (200 U) |
200 Units |
E2018-200 |
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| dsDNA Shearase™ Plus (50 U) w/ DCC™-5 (D4013) |
50 Units / 50 Preps. |
E2019-50 |
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| dsDNA Shearase™ Plus (200 U) w/ DCC™-5 (D4014) |
200 Units / 200 Preps. |
E2019-200 |
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关于5-hmC
5-hmC是近年来在动物组织中发现的，由胞嘧啶修饰而来。5-hmC在表观遗传学上的功能可能与5-甲基化胞嘧啶(5-mC)不同。尽管到现在为止还不确知其功能，有研究者猜测它在调控基因的表达与关闭过程中起着重要的作用。5-mC的发现让我们不得不重新评估DNA甲基信息，也不得不监测人类的健康组织和病理组织之间5-mC相对分布的差异。在EPI公司的MethylFlash技术之前，我们还没有发现任何直接的常规方法来检测5- hmC,以及区分5-hmC和5-mC
5-hmC 和 5-mC的区别
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