DFS PlusTaq Polymerase: more sensitivity; more processivity

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DFS-Plus Taq Polymerase:Order/request by E-mail:
DFS-Plus Taq DNA Polymerase: Datasheet
DFS-Plus Taq DNA Polymerase: Deutsche Beschreibung

Features:
DFS-Plus Taq DNA Polymerase provides a new formula in buffers and additives to prevent failures in PCR-applications were inhibitors (e.g. proteins, fat or PS) reduce the performance.
The robust enzyme is well suited for sensitive experiments using random primers or bacterial templates. Because of the high sensitivity less than 6 molecules can be detected.

Application:
Instead of conventionally purified Taq-DNA Polymerase for sensitive PCR reactions, for the detection of bacterial DNA or for applications where inhibitors decrease the performance of regular polymerases

Reaction conditions:
Same as for conventionally purified Taq-DNA Polymerase.

Concentration
5 units/μl supplied in 10 mM KPO₄ (pH 7.4 at 25°C), 0.1 mM EDTA, 0.1% Tween 20, 0.1% Triton-X 100 and 50 % (v/v) glycerol.

Unit Definition
One unit is defined as the amount of enzyme required to incorporate 10 nmoles of dNTP into acid-insoluble material in 30 min at 74°C.

Reaction Buffers provided:
Ammonium-Reaction buffer (10X)” incomplete” 
Ammonium-Reaction buffer (10X) “complete” with 25mM MgCl2
MgCl2 (100 mM)

Quality Control:
- Endonucleases Incubation of 20 units of the enzyme in 1x reaction buffer with 1 μg lambda DNA for 16 h
  at 65°C in 50 μl yields no detectable degradation of DNA.
- Incubation of 20 units of the enzyme in 1x reaction buffer with 1 μg lambda DNA EcoR I/Hind III fragments
  for 16 h at 65°C in 50 μl yields no detectable degradation of DNA.
- Incubation of 32 units of the enzyme in 1x reaction buffer with 1 μg supercoiled pUC18 DNA for 16 h at 70°
  C in 50 μl resulted in no relaxation.
- Priming activity Incubation of 40 units of the enzyme in 1x reaction buffer with 100 ng template DNA and  
  0.2 mM dNTPs each, but without primers in 100 μl resulted in no DNA synthesis.
- PCR Test Good performance of DNA amplification was confirmed by using Lambda DNA as template
  (amplified fragment 12 kb) and human placenta DNA as template (amplified fragment 3.0 kb).
- No DNA contamination with enterobacterial DNA

Transport: Shipping at ambient temperature has no negative effects on the performance
of this enzyme.

Storage: at –20 °C is recommended to safeguard against growth of bacteria
that may be introduced during handling.

Components	Volume per reaction
10X reaction buffer II	5 µl
25 mM MgCl2	1.5 µl (if necessary)
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
DFS-PlusTaq DNA Polymerase (5 u/µl)	0.1 - 0.8 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:

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