AccuPower® HotStart PCR PreMix(with UDG)


AccuPower® Hotstart PCR PreMix(with UDG) is a ready-to-use master mix for high-specificity DNA amplification, containing UDG to prevent carryover contamination or crossover contamination

The polymerase chain reaction (PCR) can amplify a single molecule over a billionfold. Thus, even minuscule amounts of a contaminant can be amplified and lead to a false positive result. Such contaminants are often products from previous PCR amplifications (carry-over contamination). Therefore, researchers have developed methods to avoid such contamination AccuPower® Hotstart PCR PreMix(with UDG) is a ready-to-use master mix for high-specificity DNA amplification, con-taining UDG to prevent carryover contamination or crossover contamination The master mix combines the Enzyme - mediated HotStart technology of HotStart DNA polymerase with integrated UDG carryover prevention technology to provide optimal performance with a variety of PCR detection technologies
 

Features and Benefits

  • Prevention of carryover contamination
UDG and dUTP in the MasterMix prevent the re-amplification of carryover PCR products between reactions. dUTP ensures that any amplified DNA will contain uracil, while UDG removes uracil residues from single- or double-stranded DNA, preventing dU-containing DNA from serving as template in future PCRs, A UDG incubation step(37°C,2min) before PCR cycling destroys any contaminating dU-containing product from previous reactions. UDG is then inactivated by the high temperatures during normal PCR cycling, thereby allowing the amplification of genuine target sequences

  • Specificity
Pyrophosphate (PPi) has high affinity for Mg2+. By adding PPi to the reaction mixture, the 2+ ions necessary for normal PCR are bound, preventing DNA polymerase activity. This PPi-Mg2+ binding prevents non-specific before PCR (zero-cycle) product formation. Upon thermal cycling, the pyrophosphatase (PPase) that is also added to the mixture is activated (>70°C) and hydrolyzes the PPi to 2 phosphate groups and facilitates the release of Mg2+, which is then available for DNA polymerase to use and resume normal activity.
  •  Easy-to-use
All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form. The user needs only to add template DNA, primers and water. Materials necessary for loading agarose gels for electrophoresis are also added in the reaction, negating the need to add loading dye after PCR is completed.
  •  Reproducibility
Each batch is produced under strict quality controls. Errors that commonly occur during mass production are eliminated during the individual packaging process. Bioneer’s current batch processing system allows for the production of more accurate and reproducible end-product yield


 

]]> Specifications

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: ~ 12 kb

 

Application

Genome template’s PCR
Low-copy target's PCR
Multiple primer pair's PCR
cDNA templates PCR
Molecular Diagnosis


Figure 1. Comparison of specificity between Accupower® HotStart PCR PreMix and HotStart PCR PreMix(with UDG).
Specificity test was operated using 7 pair of primers targeting P53 gene. Reaction mixture was incubated at 37°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 40sec at 55°C, 1min at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The amount of DNA(human) used to test is 10ng and size of PCR amplicon size each lane is represented below.
Lane 1: 139bp Lane 2: 211bp
Lane 3: 447bp Lane 4: 618bp
Lane 5: 1082bp Lane 6: 1296bp
Lane 7: 1561bp
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)


Figure 2. Comparison of sensitivity between Accupower® HotStart PCR PreMix and HotStart PCR PreMix(with UDG).
Sensitivity test was operated using serial diluted human genomic DNA. Reaction mixture was incubated at 37°C for 2min followed by, 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The amount of DNA used to test is represented below.
Lane 1: 10ng of Human genomic DNA Lane 2: 1ng of Human genomic DNA
Lane 3: 100pg of Human genomic DNA Lane 4: 10pg of Human genomic DNA
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)


Figure 3. Comparison of amplification quality using PCR products(not including uracil base) between Accupower® HotStart PCR PreMix and Accupower® HotStart PCR PreMix(with UDG).
Amplification quality test was operated using serial diluted PCR products not including uracil base. Accupower® HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37/7°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The copy number of PCR products used to test are represented below.
Lane 1: 1011copy Lane 2: 1010copy Lane 3: 109copy
Lane 4: 108copy Lane 5: 107copy Lane 6: 106copy
Lane 7: 105copy Lane 8: 104copy Lane N: No template control
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)


Figure 4. Efficiency of uracil DNA glycosylase using PCR product(including uracil base).
Efficiency test of uracil DNA glycosylase was operated using serial diluted PCR products including uracil base. Accupower® HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.
The copy number of PCR products used to test are represented below.
Lane 1: 1011copy Lane 2: 1010copy Lane 3: 109copy
Lane 4: 108copy Lane 5: 107copy Lane 6: 106copy
Lane 7: 105copy Lane 8: 104copy Lane N: No template control
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)

 


 

]]> Manual

• AccuPower® Hotstart PCR PreMix(with UDG)


MSDS
• AccuPower® Hotstart PCR PreMix(with UDG)

  
Brochure
• AccuPower® PreMix series - 2010 Brochure


Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

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