Catalogue Numbers:
BIO-21091 250 Units
BIO-21092 500 Units

Features
         • Easy visual recognition
         • Direct loading onto agarose gels
         • Same high fidelity and robust activity as ACCUZYME™ DNA Polymerase
         • Easy visual recognition
         • Amplifies fragments up to 5Kb

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

Description
ACCUZYME ™ Red DNA polymerase is a formulation of our regular ACCUZYME ™, which contains a non-toxic and non-hazardous red dye. The red dye provides easy and quick identification of reactions to which the enzyme has been added, and facilitates the confirmation of complete mixing.
When the reaction is complete, a sample of the reaction mix can be loaded directly onto the agarose gel without the need for loading buffer, since the mix is of sufficiently high density to sink to the bottom of the well. The red dye migrates towards the positive electrode, thereby providing a means to monitor the progress of the electrophoresis.
The presence of the dye has no effect on routine enzymatic manipulations, although rare exceptions may occur. To produce a reaction of sufficient density to allow for the direct loading of a sample onto a gel we recommend using a minimum of 2.5 units per 50µl reaction.

Reaction Conditions (for a 50µl volume)
10x AccuBuffer                                5µl
50mM MgCl2 Solution                   0 – 2µl
100mM dNTP Mix (see below)     0.5 – 1µl
Template and primers                    as required
ACCUZYME Red                         1 – 3µl
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 the inherent 3'-5' exonuclease activity of ACCUZYME Red, the enzyme must be added last to a reaction in order to prevent primer damage.

General Considerations:
Specificity and performance of ACCUZYME™ Red can be further improved with the use of 2x PolyMate Additive (not supplied, see associated products), which is designed for GC or AT-rich DNA, “dirty” templates or sequences with a high level of secondary structure.
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 a final concentration of 2mM Mg2+. For initial tests, use no less than 2.5 units of ACCUZYME™ Red in a 50µl reaction.
This data is intended for use as a guide only; conditions will vary from reaction to reaction and may need optimization.

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

Components:
ACCUZYME Red DNA Polymerase    250Units       500Units
ACCUZYME Red 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 Red can be stored for 12 months at -20°C.
Shipping Conditions:
On Dry Ice or Blue Ice
Unit definition:
One unit will incorporate 10nmoles of dNTPs in 30min at 72°C.

Associated Products:
Product Name                               Pack Size            Cat No
dNTP Set                                    4 x 25µmol      BIO-39025
dNTP Mix                                    1 x 500µl       BIO-39028
2x PolyMate Additive                    2 x 1.2ml        BIO-37041
ACCUZYME DNA Polymerase    250 Units       BIO-21051
SureClean Plus                               1 x 5ml            BIO-37047