Hot-StartFeatures
        • Outstanding and robust performance
        • For PCR assays requiring hot-start
        • Excellent yield in quantitative assays
        • Convenient set up at room temperature
        • Leaves ‘A’ overhang
        • Available in ready-to-go versions ImmoMix™ and ImmoMix™ Red

Applications
            • Highly suited to real-time assays
            • Products suitable for TA cloning

Description
IMMOLASE™ is a heat-activated thermostable DNA polymerase isolated from a novel organism. IMMOLASE provides improved specificity as compared to standard polymerases and can eliminate the presence of non-specifics, such as primer-dimers and mis-primed products. IMMOLASE is inactive at room temperature and therefore, prior to PCR cycling, requires activation by heat treatment for 10 minutes. Subsequently, the reaction can be handled according to the user’s existing protocols for thermostable DNA polymerases.
Specificity and performance of IMMOLASE 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.

Reaction Conditions (for a 50µl reaction)
Bioline 100 mM dNTP Mix is available as a separate product (see associated products)
Activate: pre-heating step at 95°C for 10 minutes
Denature: 94-96°C
Elongate: 72°C (allowing 15-30 seconds/kb)
This data is intended for use as a guide only; conditions will vary from reaction to reaction and may need optimization.

General Considerations:
The enzyme must be activated by heat treatment before PCR cycling. All reaction components (including IMMOLASE™) should be added to the reaction, and then pre-incubated at 95°C for 10 minutes. Subsequently, the reaction can be treated according to the user’s existing protocols for thermostable DNA polymerases.
The ideal MgCl2 concentration in the reaction is likely to be 1.5-2.5mM (final concentration), but some optimization may be necessary to achieve the best possible results. For first tests, use no less than 1 unit of IMMOLASE™ in a 50µl reaction.

    Reaction Conditions (for a 50µl reaction)

10x ImmoBuffer                           5µl
50mM MgCl2                             1.5 – 4µl
100mM dNTP Mix (see below)   0.5 – 1µl
Template and Primers                  as required
IMMOLASE™                          0.2 – 1µl
Water (ddH2O)                          up to 50µl

Components:
                                                   250 Units     500 Units     5000 Units
IMMOLASE DNA Polymerase     50µl           100µl          10 x 100µl
10x ImmoBuffer                             1.2ml        2 x 1.2ml       20 x 1.2ml
50mM MgCl2 Solution                  1.2ml           1.2ml          10 x 1.2ml

Reagent Specifications:

10x ImmoBuffer:
Separate MgCl 2 solution: 50mM MgCl2

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

Shipping Conditions:
On Dry Ice or Blue Ice

Storage and Dilution Buffer:
20mM Tris-HCl, pH 7.5, 100mM NaCl, 0.1mM EDTA, 2mM DTT, 50% Glycerol, and stabilizers.

Associated Activities:
Endonuclease and exonuclease activities were not detectable after 4 hours of incubation of 1µg of pBR322 plasmid DNA and 0.5µg Hind III-digested lambda phage DNA at 72°C in the presence of 20u of IMMOLASE.

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
ImmoMix               100 Reactions     BIO-25019
HyperLadder I          200 Lanes        BIO-33025
Agarose                       100g             BIO-41026