ThermolaseTA™-Thermoactivated DNA polymerase.
Description
ThermolaseTA™ (EC 2.7.7.7) is a thermostable DNA-dependent DNA polymerase that is inactive until incubation at 95°C for 15 min. During thermoactivation step the nonspecific primer annealing is prevented that results in increase of an amplification specificity.
Unit definition

One unit of the enzyme will incorporate 10 nmol of dNTPs into an acid-insoluble form in 30 min at 72°C under standard assay conditions using a DNA template.

Reaction buffer
B35 (x10, with Mg2+ ): 100 mM Tris-HCl, pH 8.3, 500 mM KCl, 25 mM MgCl2.

Amplification conditions

For amplification of 1,5 kb fragment use 0,2 ml tubes and following conditions:
10x reaction buffer
2 mM dNTPs (200 µM each)
human genomic DNA (100 ng)
forward primer factor VIII.3(25 pM)
reverse primer factor VIII.4 (25 pM)
Thermolase™TA (5 units/µl)
H2O
 
 
3 µl
3 µl
1 µl
1 µl
1 µl
0,5 µl
20,5 µl

total

30 µl

Hot-start

Steps

95° 15 min
 
58° 0,5 min
72° 2 min
93° 15 sec

Number of cycles

30 cycles

 

Storage
Store at -20°C.
Applications
hot-start PCR.

Quality control

activity, SDS-PAGE purity, absence of endo- and exonucleases, specific performance tests.

Catalog #
T2.4

T2.5

 

 

 

Pack size

1000 U

2500 U

 

 

 

 

Copyright © 2002 M.B. Enzymes GmbH