( re 1A) or SureStart Taq (Fig...)Greater Amplification Specificity with New Hot Start PCR Enzyme

re 1A) or SureStart Taq (Figure 1B) DNA polymerase. Taq DNA polymerase amplified numerous extraneous bands in addition to the desired product (Figure 1A, Lanes 1-5). In contrast, background was dramatically reduced in reactions carried out with SureStart Taq DNA polymerase (Figure 1B). High product yields were achieved after 30 to 33 cycles when SureStart Taq was activated prior to temperature cycling during a 10-minute incubation step (95C) added at the beginning of the thermal cycling program (Figure 1B, Lanes 1-5). When the pre-PCR heat step was omitted, 36 to 39 cycles were required to achieve comparable product yields (Figure 1B, Lanes 6-10). Although the PCR heat activation method generally requires more cycles (35 to 45 cycles) to achieve optimal product yield, greater amplification specificity may be realized due to lower enzyme concentrations in initial PCR cycles.

Superior PCR Performance

Amplification of low-copy-number templates is especially problematic with complex DNA samples due to frequent mispriming events at ambient temperature. Hot start methods eliminate mispriming and thereby improve the detection of low-copy-number targets. Challenging model amplification systems were used to evaluate the hot start properties of SureStart Taq DNA polymerase. In the systems employed in Figures 2 and 3, amplicons were synthesized from serially diluted DNA templates in the presence of high concentrations of denatured genomic DNA. In Figure 2, a 115-bp fragment of the gag gene was amplified from 5 to 5000 copies of HIV-1 DNA in the presence of 0.1 mg of denatured human genomic DNA. While Taq DNA polymerase successfully amplified t
'"/>

Source:

Page: All 1 2 3 4 5 6 7
Related biology technology :

1. Enjoy a Greater Sense of Security with PCR-Grade Water
2. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
3. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
4. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
5. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
6. Improve Amplification Specificity with Hot Start PCR Enzyme
7. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
8. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
9. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
10. Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler
11. Amplification of Epstein-Barr Virus Exon C with Difficult Primers Using Eppendorf Mastercycler gradient