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An Outstanding Robust Polymerase for Hotstart PCR assays, highly suited for real time Assays with PCR products suitable for TA cloning
Applications:
The product is highly suited for real time assays. The resulting PCR Products are suitable for TA cloning
Description- AuPreP Hotstart TaQ DNA Polymerase is a heat-activated thermostable DNA polymerase isolated from a novel organism. It provides improved specificity as compared to standard polymerases and can eliminate the presence of non-specifics, such as primer-dimers and mis-primed products. The enzyme 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.
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Reaction Conditions (for a 50μl reaction)
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| 10x Buffer |
5μl |
| 50mM MgCl 2 |
1.5 – 4μl |
| 100mM dNTP Mix (see below) |
0.5 – 1μl |
| Template and Primers |
as required |
| AuPreP Hotstart DNA Polymerase |
0.2 – 1μl |
| Water (ddH 2 O) |
up to 50μl |
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Activate: pre-heating step at 95°C for 10 minutes
Denature: 94-96°C
Elongate: 72°C (allowing 15-30 seconds/kb)
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We recommend to use PCR DOCTOR for GC / AT / Dirty / Difficult Templates. Hence order it separately if needed. How to use the PCR
DOCTOR ? Compose the reaction mix, containing buffer, dNTPs, Mg 2+ , template DNA, primers, DNA polymerase. Add 2x PCR DOCTOR at the volume of half of the final volume of the reaction (e.g. 25ul per 50ul final volume, etc). Add ddH 2 O up to the final volume and mix with pipetting
This data is intended for use as a guide only; conditions will vary from reaction to reaction and may need optimization.
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General Considerations:
The enzyme must be activated by heat treatment before PCR cycling. All reaction components (including AuPreP Hotstart DNA Polymerase) 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 MgCl 2 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 AuPreP Hotstart DNA Polymerase in a 50μl reaction.
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Components: |
| 250 Units |
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500 Units |
| AuPreP Hotstart DNA Polymerase |
50μl |
100μl |
| 10x Buffer |
1.2ml |
2 x 1.2ml |
| 50mM MgCl 2 Solution |
1.2ml |
1.2ml |
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Storage Conditions: @ -20°C for 1 year
Shipping Conditions: Dry Ice/ Blue Ice
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Storage and Dilution Buffer:
20mM Tris-HCl, pH 7.5, 100mM NaCl, 0.1mM EDTA, 2mM DTT, 50% Glycerol, and stabilizers.
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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 AuPreP Hotstart DNA Polymerase.
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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. |
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| RELATED PRODUCTS |
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AuPreP Oligos (Oligo synthesis available in different scales, purifications & modifications)
AuPreP TaQ DNA Polymerase ( Ultrapure, Ultra-stable & Ultra-sensitive Taq DNA Polymerase)
AuPreP Hotstart TaQ DNA Polymerase (Robust Polymerase for Hotstart PCR assays)
AuPreP Super Fidelity TaQ DNA Polymerase (High fidelity Polymerase produces blunt ended amplicons upto 5Kb)
PCR Doctor - (PCR enhancement additive used with AuPreP Hotstart Taq or AuPreP Super Fidelity Taq
especially designed for GC/AT/Dirty/Difficult Templates
AuPreP Red PCR Master Mix ( 2x Master mix with Red Dye without Enhancer)
AuPreP DIAMOND MASTER-MIX (2x Mastermix with PCR Enhancer & Stabilizer but without dye)
AuPreP DIAMOND DOUBLE DYE MASTERMIX (2x Mastermix with PCR Enhancer, Stabilizer & tracking dyes)
AuPreP Gold cDNA Synthesis Kit (cDNA Synthesis Kit using RT with reduce Rnase H activity)
AuPreP Gold RT-PCR Combo Kit ( two-step RT-PCR protocol with tracking dye )
Novascript III Rnase H‾ RT (Premium Rnsase H‾ Reverse transcriptase offering top performance)
Novascript III single step RT-PCR System (1step RT-PCR system using Novascript & AuPreP Hotstart DNA Polymerase) |
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