Hot start functionality

This "hot start" product utilizes a Taq-specific monoclonal antibody to minimize mis-priming and/or formation of primer-dimer before thermal cycling.

• Complete:
  OmniMix™ HS contains all of the necessary reagents for PCR, except primers and probe or intercalating dye.
  
• Fast:
  No additional time required for activation.
  
• Specific:
  Reduction of nonspecific product can result in improved detection.

Easy to use format

The ready-to-use format of OmniMix HS simplifies assay preparation while adding flexibility. Larger quantities can be made by using multiple beads.

• Simplified:
  Eliminating extra pipetting steps and multiple reagents reduces contamination risk and provides better reproducibility.
  
• Stable:
  OmniMix HS is a lyophilized bead that is stable at 2-8 °C.
  
• Scaleable:
  Each 0.5 mL tube contains an optimized formula of reagents for performing two 25 µL PCR amplification reactions.

Making reproducible PCR easy to perform:

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Consistency of results with OmniMix HS

Comparison of liquid reagents, individual OmniMix HS beads, and pooled OmniMix HS beads. Cycle threshold and endpoint fluorescence values obtained with a TaqMan assay incorporating a FAM-labeled probe. 100 fg of input DNA represents the limit of detection for this assay. A p value of greater than 0.05 was obtained indicating no significant difference among the three data sets.

	Liquid Bulk	Pooled Beads	Individual Beads
Mean CT	32.9	32.2	32.5
SD	0.3	0.3	0.3
% CV	0.9	0.8	1.1
Mean End Point Fluorescence	251.8	220.8	281.7
SD	34.6	38.6	37.9
% CV	13.7	17.5	13.4
N=	96	96	96

Hot start performance

Experimental results obtained with and without the use of hot start Taq polymerase are shown below. Neisseria gonorrhoea assay with intercalating dye and TaKaRa HS Taq polymerase or a non-hot start polymerase in liquid reagent format. Data represents the first derivative peaks from melt curve analysis immediately following 45 cycles of PCR in the presence of 0.5X intercalating dye.

Negative control: non-hot start vs hot start.

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Negative control: non-hot start vs hot start.
300 molecules: non-hot start vs hot start.

Top graph shows experimental results obtained without the use of hot start Taq polymerase. In ten out of ten replicates non-specific product was generated with a Tm of ~75 and ~80.

Bottom graph shows experimental results obtained with the use of hot start Taq polymerase. An 80% reduction of non-specific product is observed (flat line). Click pictures for larger images.

300 molecules: non-hot start vs hot start.

Top graph shows experimental results obtained without the use of hot start Taq polymerase. Ten replicates show a mixed population of non-specific product (Tm ~ 80) and specific product (Tm ~85). The melt analysis indicates a greater proportion of non-specific to specific products generated.

Bottom graph shows that with the use of hot start Taq polymerase only specific product is generated (Tm ~85).

Content

When reconstituted to a final volume of 50 µL, OmniMix HS contains:
3U TaKaRa hot start Taq polymerase
200 µM dNTP
4 mM MgCl2
25 mM HEPES buffer pH 8.0± 0.1

Packaged for by Cepheid