EheI (NarI*)
Available in FastDigest® format
* Unlike NarI, EheI produces fragments
with blunt ends.
digested lambda DNA
0.7% agarose 5'-G G C^G C C-3' 3'-C C G^C G G-5'#ER0441
Supplied with:
10X Buffer Tango™500 u
1 mlCommercial Isoschizomers:
(BbeI), DinI, EgeI, (KasIm), (Mly113I), (NarIm), SfoIm
Enzymes in parentheses have different cleavage specificities.
"m" indicates different sensitivity to methylation.Related Documents (in pdf, ~55 KB):
Certificate of Analysis: #ER0441
MSDS (English)
MSDS (English-USA)
MSDS (German)
Chart (in pdf, 721 KB)
Brochure (in pdf, 316 KB)Concentration
10 u/µlConditions for 100% Activity
Recommended
bufferIncubation
temperatureUnits for
overnight
incubation,
u/µg DNAThermal
inactivation,
in 20 minEnzyme activity, %
B
(blue)G
(green)O
(orange)R
(red)Tango™
(yellow)1X 1X 1X 1X 1X 2X Buffer Tango™ 37°C 1.0 65°C 20-50 50-100 0-20 0-20 100 20-50 Storage Buffer
10 mM Tris-HCl (pH 7.5 at 25°C), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.Ligation and Recleavage
After 50-fold overdigestion with EheI, more than 80% of the digested pBR322 DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1 µg of fragments and 10% PEG. More than 90% of these can be recut.
Methylation Effects Dam:
Dcm:
CpG:
EcoKI:
EcoBI:never overlaps
may overlap
completely overlaps
never overlaps
never overlaps- no effect.
- no effect.
- blocked.
- no effect.
- no effect.Digestion of Agarose-embedded DNA
Minimum 20 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Note
- Unlike NarI, EheI completely digests lambda and pBR322 DNAs.
- Low salt, high glycerol (>5%) concentrations, pH >8.0 or large excess of the enzyme may result in star activity.
Double Digestions using Tango™ Buffer, DoubleDigest™
Optimal
bufferEnzyme activity, %
Tango™
(yellow)1X 2X Buffer Tango™ 100 20-50 Number of Recognition Sites in DNA Molecules
Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184 1 2 1 4 1 1 0 0 0 0 4
See also General Properties of Restriction Enzymes:
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Updated balandžio 22, 2008 11:20