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- 文献和实验
- 技术资料
- 英文名:
DpnI (10 U/µL)
- 规格:
500 units
描述
| 5' | G | Am6↓ | T | C | 3' | ||||
| 3' | C | T ↑ | Am6 | G | 5' |
Thermo Scientific DpnI restriction enzyme recognizes Gm6A^TC sites and cuts best at 37°C in Tango buffer (isoschizomers: MalI). See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion.
Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.
Features
• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities
规格
| Compatible Buffer: | 10x Buffer Tango |
|---|---|
| Enzyme: | Dpn I |
| Methylation Sensitivity: | Not CpG methylation-sensitive, Not dcm methylation-sensitive, dam methylation-sensitive |
| Optimal Reaction Temperature: | 37° C |
| Product Size: | 500 units |
| Sensitive to Heat Inactivation: | Yes |
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文献和实验DpnI mediated site-directed Mutagenesis
, 68°C 2 minutes/kb of plasmid length 3. Degradation of methylated (parental) DNA with DpnI Cool down PCR reaction. Add 1µl Dpn I (10 unit) to PCR reaction 37°C and incubate for 1 hr. 4. Transformation into E. coli
DpnI mediated site-directed Mutagenesis
length 3. Degradation of methylated (parental) DNA with DpnI Cool down PCR reaction. Add 1µl Dpn I (10 unit) to PCR reaction 37°C and incubate for 1 hr. 4. Transformation into E. coli Place 200 µl highly competent cells (1 x 108/ug efficiency
ng) 1ulprimer 1 (125ng) 1ulprimer 2 (125ng) 1ulpyrobest DNA polymerase(TaKaRa)(5U/ul) 0.25ul加无菌蒸馏水至 50ul 3、产物沉淀纯化:加1/10 体积的醋酸钠,1倍体积的异丙醇,混匀置冰上(或-20゜C冰箱)5min,离心弃上清,70~75%乙醇洗盐两次
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