Need a quotation? Create it here!
No login/account required.
No login/account required.
Your Basket/Online Quote
Items: 0 (0,00 €)
» Search & Order
» Sign in / Register
Your Basket/Online Quote
Items: 0 (0,00 €)
» Search & Order
» Sign in / Register
Neoschizomers: BfuCI, Bsp143I, BstENII, BstKTI,BstMBI, DpnII, Kzo9I, NdeII
JBSpeed Restriction Enzyme
| Cat. No. | Amount | Price (EUR) | Buy / Note |
|---|---|---|---|
| EN-160S | 200 Units | 39,70 | Add to Basket/Quote Add to Notepad |
| EN-160L | 5 x 200 Units | 158,80 | Add to Basket/Quote Add to Notepad |
For general laboratory use.
Unit Definition: One unit is the amount of enzyme required to completely digest 1 μg of pBR322 (22 sites) in 1 hour in a total reaction volume of 50 μl. Enzyme activity was determined in the recommended reaction buffer.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid (Supplied in 10 mM Tris-HCl pH 7.4, 400 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA and 50 % [v/v] glycerol)
Concentration: 10 units/μl
Source: Diplococcus pneumoniae G41, recombinant, E. coli
Supplied with: 10x Universal Buffer (UB)
Recommended 50 μl assay
| 5 μl | 10x Universal Buffer (UB) |
| 1 μg | pure DNA1 or PCR product2 |
| 10 units | enzyme |
| fill up to 50 μl | PCR grade water |
Protocol:
Double Digestion - Buffer Compatibility:
B1 - 75-100 % Relative Activity
B2 - 75-100 % Relative Activity
B3 - 50-75 % Relative Activity
B4 - 10 % Relative Activity
B5 - 75-100 % Relative Activity
1x UB - 100 % Relative Activity (recommended)
Please note that the optimum digestion condition for this enzyme is 1x UB. Within the Universal Buffer (UB) system, the most majority of our enzymes display 100% Relative Activity in 1x UB and only few either in 0.5x or 2x UB. If optimum condition for second enzyme is different than the recommended for the first enzyme, we suggest carrying out first the restriction at the higher recommended concentration of UB and dilute the reaction volume to the adequate UB concentration for further proceeding with the second restriction.
Reaction Enzymes Buffer Guide:
| Buffer 1 | 10 × B1 | 100 mM 100 mM 1000 μg/ml | Tris-HCl (pH 7.9, 25°C) MgCl2 BSA |
| Buffer 2 | 10 × B2 | 100 mM 100 mM 500 mM 1000 μg/ml | Tris-HCl (pH 7.9, 25°C) MgCl2 NaCl BSA |
| Buffer 3 | 10 × B3 | 500 mM 100 mM 1000 mM 1000 μg/ml | Tris-HCl (pH 7.9, 25°C) MgCl2 NaCl BSA |
| Buffer 4 | 10 × B4 | 100 mM 100 mM 1500 mM 1000 μg/ml | Tris-HCl (pH 7.9, 25°C) MgCl2 NaCl BSA |
| Buffer 5 | 10 × B5 | 200 mM 100 mM 500 mM 1000 μg/ml | Tris-acetate (pH 7.9, 25°C) Mg-acetate K-acetate BSA |
Reaction Buffer Compatibility:
Our restriction enzymes are fully compatible to restrictases and buffer systems from other manufacturers and can be used along in double digestions. To obtain best results, consult the corresponding manuals of all involved products.
Ligation and recutting:
After 50-fold overdigestion with DpnI, >70 % of the DNA fragments can be ligated and >95 % of these can be recut.
Methylation dependency:
DpnI is specific for methylated and hemimethylated DNA. Since DNA isolated from most E. coli strains is dam methylated, it is susceptible to DpnI digestion. Hence, DpnI is frequently used after a PCR reaction to digest the methylated parental DNA template and select for the newly synthesized DNA containing mutations.
Quality Control:
All preparations are assayed for contaminating endonuclease, 3'-exonuclease, 5' exonuclease/ 5' phosphatase, as well as nonspecific single- and doublestranded DNase activities.
Related products: Universal Restriction Buffer, #EN-300
BIOZ Product Citations:
Powered by Bioz
