TspDT I

TspDT I Source

Purified from E coli strain that carries the cloned tspDTRI gene from Thermus species DT

Unit Definition

One unit is the amount of enzyme required to completely digest 1 µg of pUC19 DNA to obtain a stable digestion pattern in 1 hour in a total reaction volume of 30 µl

Star Activity

N/A

Reaction Temperature

70°C

Heat Inactivation

No

Assay Unit Substrate

pUC19 DNA

Reaction Buffer

10 mM Tris-HCl (pH 8.5 at 25°C)
10 mM MgCl₂
1 mM dithiothreitol
+Enhancers (1)

Notes:

1. Avoid multiple cycles of freezing / thawing of the stock reaction buffer; No more than three [3] times.
2. Thawing should be preformed at temperatures not exceeding 10°C.
3. Recommended procedure is to divide the provided reaction buffer into smaller portions and preserve them at -70°C for long term storage.
4. Recommended short term storage of reaction buffer is at         -20°C.

Storage Buffer

20 mM Tris-HCl (pH 8.3 at 25°C)
25 mM (NH₄)₂SO₄ 
25 mM KCl
0.5 mM EDTA
0.5 mM dithiothreitol
0.02% Triton X-100™
0.02% Tween 20™
0.02% Igepal
50% (v/v) glycerol

Assay Conditions 

10 mM Tris-HCl (pH 8.5 at 25°C)
10 mM MgCl₂ 
1 mM dithiothreitol 
1 µg pUC19 DNA
+Enhancers (1)
Incubation is at 70°C for 1 hour in a reaction volume of 30 µl 

Notes

1. It is not recommended to use more than 2 units per 30 µl reaction. It is suggested to perform digestion for over 1 hour.
2. To avoid DNA shift during electrophoresis caused by strong protein-DNA interaction, it is recommended to terminate reaction by addition of reaction stop solution (containing denaturing reagent, i.e. 0.2% SDS) followed by heat inactivation at 89°C for 20 minutes.

Storage Conditions

Store at -20°C
Shipped on dry ice

Categories

Restriction Endonucleases