HindIII is a restriction enzyme purified from a recombinant E.coli strain.
Reagents supplied: 10x R2 and 10x EQ buffer
Unit definition: One unit is the amount of enzyme required to digest 1 μg of Lambda DNA in 60 min at 37oC in a total reaction volume of 50 μl.
Standard reaction conditions: In order to overcome DNA source variability, a ~10 fold overdigest is suggested:
10U Hind III
10x EQ buffer
Sterile ultrapure water
Up to 50 μl
Speed Digest reaction: Incubate for 15 minutes as in standard reaction in 1x R2 buffer (50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25oC), 10 mM MgCl2, 100 μg/ml BSA) or 1x eQ buffer (20 mM Tris-acetate (pH 7.9 @ 25oC), 10 mM Mg-acetate, 50 mM K-acetate, 100 μg/ml BSA).
Activity(%) in EnzyQuest buffers-Double digest tips
Reactions were performed as for unit definition reaction. The above data could be used as guideline for double digest reactions. Our enzymes are also active and compatible in buffer systems from other vendors.
Heat inactivation: 65oC for 20 minutes
dam methylation: Not sensitive
dcm methylation: Not sensitive
CpG methylation: Not sensitive
Unit calculation assay conditions:
1x R2 buffer
Incubation at 37oC
50 mM KCl,
10 mM Tris-HCl (pH 7.4)
0.1 mM EDTA
1 mM dithiothreitol
500 μg/ml BSA
Store at -20oC
Non-specific nuclease activity: 700 units of Hind III do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC.
Re-ligation and recut: After 100-fold overdigestion with Hind III, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.
Star activity: Star activity may be observed in the presence of Mn2+.
Strictly Necessary Cookies
Strictly Necessary Cookie should be enabled at all times so that we can save your preferences for cookie settings.
If you disable this cookie, we will not be able to save your preferences. This means that every time you visit this website you will need to enable or disable cookies again.
3rd Party Cookies
This website uses Google Analytics to collect anonymous information such as the number of visitors to the site, and the most popular pages.
Keeping this cookie enabled helps us to improve our website.
Please enable Strictly Necessary Cookies first so that we can save your preferences!