Catalogue Numbers:
BIO-21051 250 Units
BIO-21052 500 Units

Features
       • High fidelity coupled with high yield
       • Amplifies fragments up to 5Kb
       • Available as a convenient pre-mixed, pre-optimized solution (ACCUZYME™ Mix)

Applications
       • Ultra-high fidelity for subsequent cloning
       • Blunt-end cloning
       • DHPLC compatible (detergent free)

Description
ACCUZYME™ is a thermostable enzyme possessing 5’-3’ DNA polymerase and 3’-5’ proofreading exonuclease activities, offering high fidelity. ACCUZYME produces blunt-ended amplicons of up to 5Kb in length.
ACCUZYME is supplied with 10x Reaction Buffer containing MgSO4, which provides optimal final reaction conditions (1.5mM Mg2+) for most experiments. In order to allow optimization of reaction conditions, additional MgCl2 is provided.
The specificity and performance of ACCUZYME can be further improved with the use of 2x PolyMate Additive
(Cat No. BIO-37041), which is designed for GC or AT-rich DNA, “dirty” templates or sequences with a high level of secondary structure.

Reaction Conditions (for a 50µl volume)
10x AccuBuffer                            5µl
50mM MgCl2 Solution                0.5 – 1µl
100mM dNTP Mix (see below)    0 – 2µl
Template and primers                  as required
Enzyme                                       1 – 3 µl
Water (ddH2O)                          up to 50µl

100mM dNTP Mix is available as a separate product (see associated products).
Denature: 94-97°C
Extension: 68°C Allowing 0.5 -1 minutes per Kb
Owing to ACCUZYME's inherent 3'-5' exonuclease activity, the enzyme must be added last to a reaction in order to prevent primer damage.
This data is intended for use as a guide only; conditions will vary from reaction to reaction and may need optimization.

General Considerations:
The suggested final concentration of Mg2+ in the reaction is likely to be 2-4mM, but some optimization may be necessary to achieve the best possible results. Please, note that the reaction buffer already provides 2mM Mg2+. For first tests, use no less than 2.5 units of ACCUZYME in a 50µl reaction.

Product Specifications
Batch details:
Batch No: See vial
Units per vial: See vial
Concentration: 2.5u/µl

Components
                                                            250 Units                500 Units
ACCUZYME DNA Polymerase            100µl                       200µl
10x AccuBuffer                                      1.2ml                    2 x 1.2ml
50mM MgCl2 Solution                           1.2ml                      1.2ml

Reagent Specifications:
10x AccuBuffer: 600mM Tris-HCl, 60mM (NH4)2SO4, 100mM KCl, 20mM MgSO4, pH 8.3 at 25°C
MgCl 2 Stock Solution: 50mM MgCl2
ACCUZYME Storage Buffer: 20mM Tris-HCl, pH 7.5, 100mM NaCl, 0.1mM EDTA, 2mM DTT, 50% Glycerol, and stabilizers.

Storage Conditions:
ACCUZYME™ can be stored for 12 months at -20°C.

Shipping Conditions:
On Dry Ice or Blue Ice

Unit definition:
One unit is defined as the amount of enzyme that incorporates 10nmoles of dNTPs into acid- insoluble form in 30 minutes at 72°C.

Associated Products:
Product Name                        Pack Size         Cat No
dNTP Set                            4 x 25µmol      BIO-39025
dNTP Mix                              500µl            BIO-39028
2x PolyMate Additive           2 x 1.2ml         BIO-37041
AccuSure DNA Polymerase  250 Units        BIO-21068
HyperLadder I                     200 Lanes        BIO-33025
Agarose                                  100g             BIO-41026

Product Citations:
1. Wolkow, T.D. & Enoch, T. BMC Genetics 4(6) (2003).
2. Chan Kwo Chion, C.K., et al. Applied and environmental microbiology 71(4), 1909-1914 (2005).
3. Mandola, M.V., et al. Cancer Research 63(11), 2898-2904 (2003).