Maximo Taq-Blue DNA Polymerase: Datasheet
Features:
Maximo Taq-Blue DNA Polymerase provides robust PCR performance in a wide range of PCR applications and different templates. After PCR reaction the enzyme can be loaded to agarose gel, no dye and DNA loading buffer is needed. The enzyme is time- and cost saving, because it includes dye and loading buffer, already.
Applications:
- Standard / General PCR with visible control
- High-throughput PCR
- Primer extension
- Gene mutation
- T/A cloning
Description:
Maximo Tag-Blue DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity and a double-stranded specific 5´→3´ exonuclease activity. The enzyme consists of a single polypeptide with a molecular weight of 94kDa.
Concentration: 1 u/µl
Unit definition:
One unit incorporates 10 nmol of deoxyribonucleotide into acid-precipitation material in 30min at 74 degree
Storage Buffer:
25mM Tris-HCl (pH8.0), 100mM KCl, 0.1mM EDTA, 1mM DTT, 50% Glycerol, 0.5% NP 40, 0.5% Tween 20, in blue loading buffer
Reaction Buffers supplied with the enzyme:
10X Buffer I: 500mM KCl, 100mM Tris-HCl, pH 9.0, 1% Triton X-100, 15mM MgCl2
10X Buffer II: 500mM KCl, 100mM Tris-HCl, p H 9.0, 1% Triton X-100
MgCl2: 25 mM
Quality control:
- PCR with various templates – genomic DNA, Phage Lambda DNA
- 2,2 kb DNA amplification from 50 ng DNA
- batch variation and level of bacterial DNA contamination
Transportation: on blue ice
Storage: at -20°C for 12 months
Usage:
|
Components |
Volume per reaction |
|
10X reaction buffer I or buffer II |
5 µl |
|
25 mM MgCl2 |
1.5 µl (if you use buffer II) |
|
dNTP-Mix (40mM) |
1.0 µl |
|
Up-stream primer (10 µM stock) |
0,5-2.5 µl |
|
Down-stream primer (10µM stock) |
0.5-2,5 µl |
|
Template DNA |
0.1-15 ng/ml plasmid DNA
1-10 µg/ml genomic DNA |
|
Maximo Taq DNA-Blue Polymerase (1 u/µl) |
0.8 µl - 2 µl |
|
Sterile dest. Water (molecular grade) |
up to 50 µl total reaction volume |
Note:
- vortex all solutions carefully before using
- dispense all reagents on ice
- add the enzyme after Template DNA
- may you have to optimize the MgCl2 concentration for best result
General Thermo-Cycler protocol:
|
Step |
Time |
Temperature |
|
Initial denaturation |
2-5 min |
94-95°C |
25-30 Cycles:
Denaturation
Annealing
Extension
|
10-25 sec
10-25 sec
60 sec |
94-95°C
55-65°C
72°C per 1kb
|
|
Final extension |
5 min |
72°C |
|
