Sequencing Pol

Taq Pol mutant for incorporation of ddNTPs

Thermus species, recombinant, E. coli

Product Cat. No. Amount Price (EUR) Buy / Note
S pack PCR-206S 200 units 70,00 Add to Basket/Quote Add to Notepad
L pack PCR-206L 1000 units 280,00 Add to Basket/Quote Add to Notepad

For in vitro use only!

Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmol of dNTP into an acid-insoluble form in 30 minutes at 74 °C.

Shipping: shipped on blue ice

Storage Conditions: store at -20 °C
avoid freeze/thaw cycles

Shelf Life: 12 months

Form: liquid

Concentration: 5 units/μl

Availability Restriction: Exclusively distributed in Japan by Greiner BioOne

Description:
Sequencing Pol is a Taq polymerase mutant showing an enhanced efficiency for incorporation of ddNTPs. Its capability of incorporating ddNTPs and dNTPs at equal rates makes it the ideal choice for DNA cycle sequencing.
The enzyme replicates DNA at 74 °C. It catalyzes the polymerization of nucleotides into duplex DNA in 5'→3' direction in the presence of magnesium and has a 5'→3' polymerisation-dependent exonuclease replacement activity only.
Kit contents:
Sequencing Pol (red cap)
5 units/μl Sequencing Polymerase in storage buffer
Sequencing Buffer without MgCl2 (green cap)
10x conc.

500 mM Tris-HCl, pH 9.5 (25 °C)
MgCl2 Stock Solution (yellow cap)
25 mM MgCl2

Recommended 50 μl PCR assay:

5 μl10x Sequencing Bufferblue cap
6 μlMgCl2 stock
solution
yellow cap
200 μMeach dNTP-
0.2 - 1 μMeach Primer-
2 - 50 ngtemplate DNA-
0.5-1 μl
(2.5-5 units)
Sequencing Polred cap
Fill up to 50 μlPCR-grade water-

Optimization of MgCl2 concentration:
A concentration of 2.5-3.5 mM Mg2+ is recommended for most applications. For an individual optimization add MgCl2 stock solution as shown in the table below.
50 μl PCR assay
MgCl2 Stock Solution5 μl6 μl7 μl8 μl
final MgCl2 conc.2.5 mM3 mM3.5 mM4 mM