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LightCycler® 1536 System Performance Data

 

Read in this article:

 

Sensitivity and Reproducibility

Superb Linear Range

Figure 1: Superb linear range of the LightCycler® 1536 System. Eight concentration steps of a 1:10 serial dilution from 107 to 100 copies/sample and negative controls (96 replicates each).

 

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Outstanding Sensitivity and Reproducibility

Figure 2: Outstanding sensitivity and reproducibility of the LightCycler® 1536 System. Sixteen concentration steps of a 1:2 serial dilution from 106 to 3 x 101 copies/sample (96 replicates each) distributed in a checkerboard scheme across the entire multiwell plate, as depicted in the crossing point heatmap chart (6-times 4 x 4 replicates).

 

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Real-Time PCR Results

Precision Melting Curve Analysis for DNA Product Identification

Figure 3: Precision melting curve analysis for DNA product identification with the LightCycler® 1536 System. Melting peak curves of two individual target genes, illustrated as a first negative derivative plot of the fluorescence versus temperature.

 

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Sensitive Detection of Endpoint Fluorescence (EPF) Values

Figure 4: Sensitive detection of endpoint fluorescence (EPF) values. Amplification curves and one corresponding heatmap showing endpoint values of a dual-color fluorescence detection analysis.

 

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Application Data

High-Throughput Gene Expression Profiling

Figure 5: High-Throughput Gene Expression Profiling.

Experimental setup:

Sixteen tissue samples were analyzed using 96 RealTime ready Assays, with 82 target genes from the apoptosis pathway, 8 reference genes, and 6 controls.

Results:

  • LightCycler® 1536 Software:
    a) Amplification data for all 1536 real-time PCR assays; b) Single analysis of 1 target gene (AKT1) in 16 tissue samples; c) Single analysis of 82 target genes in 1 tissue sample (small intestine)
  • Downstream software:
    connectionParserd) Normalized ratios of 1 target gene (AKT1) in 15 tissue samples; e) Normalized ratios of 82 target genes in 1 tissue sample (small intestine)

 

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Long-Term Stability

Figure 6: Long-term stability of the RealTime ready DNA Probes Master. Tenfold dilution steps of cDNA (starting material: 500 and 50 pg total RNA equivalents) were assayed either immediately after PCR setup or after 8, 24, and 48 hours at room temperature (standard deviation: 500 pg total RNA: 0.14; 50 pg total RNA: 0.15).


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