Unit definition
One unit is the amount of enzyme required to catalyze the incorporation of 10 nanomoles of nucleotides into acid insoluble material in 30 min.
at 74C under assay conditions.
Storage conditions
20mM Tris-HCl pH 8.0, 100mM KCl,
0.1mM EDTA, 1mM dithiothreitol,
0.5% Tween 20, 0.5% Nonidet P40,
50% glycerol. Stored at -20° C.
Incubation buffer with MgCL2(1X)
10mM Tris-HCl pH 9.0, 50mM KCl,
1.5mM MgCl2, 0.1% Triton X-100,
0.2 mg/ml BSA
Incubation buffer w/o MgCL2(1X)
10mM Tris-HCl pH 9.0, 50mM KCl,
0.1% Triton X-100, 0.2 mg/ml BSA |
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Figure a:
Amplification of a 400 bp fragment of human ß globin with Taq DNA polymerase. Varying amounts of human ß globin, 1 pg to 100 ng,
were amplified under the following conditions: 50 µl reaction; 50 pmoles each specific primer, 300 µM each dNTP, 1U Taq DNA pol, MgCl2
1.5 mM, 5 min at 93°C then 1 min at 91°C, 1 min at 62°C, 2 min at 70°C for 37 cycles, 1 min at 91°C, 1 min at 62°C and 10 min at 70°C
for 1 cycle.Lane 1: marker pBR322/HaeIII, Lane 2: 1pg, Lane 3: 25 pg, Lane 4: 250 pg, Lane 5: 1ng, Lane 6: 10 ng, Lane 7: 100 ng, of template DNA respectively. 
Figure b:
Amplification of a 1,600 bp DNA insert in pUC with Taq DNA polymerase. Varying amounts of pUC containing a cloned insert 1 pg to 1
ng were amplified under the following conditions: 50 µl reaction; 50 pmoles each specific oligo, 300 µM each dNTP, 1U Taq DNA pol,
MgCl2 1.5 mM, 30 sec at 94°C, 1 min at 68°C, 2 min at 70°C for 37 cycles, 1 min at 94°C, 1’ at 68°C and 10 min at 70°C for 1 cycle.
Lane 1: 1pg, Lane 2: 10pg, Lane 3: 100pg, Lane 4: 1ng, of template DNA respectively. |