Ahram Biosystems Inc.

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Overview
Palm PCR device is loaded with fully validated operation protocols that enable ultra-fast nucleic acid amplification with high sensitivity. In each protocol operation, selection of the PCR speed level makes the Palm PCR device perform PCR amplification at the pre-defined speed with the dynamic range listed in the Palm PCR Selection Chart. A few examples are described below for the high performance of the Palm PCR system.

Contents

Ultra-Fast Amplification
< Example 1: Amplification in the F1 speed level >
Shown in this first example are the results of amplifying 510 bp β-globin from human genomic DNA in the F1 speed level. The PCR reaction volume was 20 μL and amount of the template DNA used was 10 ng, corresponding to 3,000 copies. PalmTaq™ High-Speed DNA polymerase (Part No. RK1010 and RK1020) was used. Speed of the thermal cycling in the F1 speed level is 1 minutes/cycle, corresponding to 30 minutes/30 cycles.

As shown, the PCR amplification reaches near saturation level within 30 minutes (30 cycles) without generating any side bands. The amplified band is sufficiently strong even in the 25 minutes PCR time, demonstrating the high speed and efficiency of the Palm PCR system.

< Example 2: Amplification in the T2 speed level >
In this example, 249 bp α-tublin was amplified from human genomic DNA in the fastest speed level T2. The PCR reaction volume was 20 μL and amount of the template DNA was 10 ng, corresponding to 3,000 copies. PalmTaq™ High-Speed DNA polymerase (Part No. RK1010 and RK1020) was used. Speed of the thermal cycling in the T2 speed level is 36 seconds/cycle, corresponding to 18 minutes/30 cycles.

As shown, the PCR amplification is completed in extremely short time with high efficiency and accuracy. Only after 15 minutes (25 cycles), the amplified band becomes significantly strong and it reaches near saturation level after 18 minutes (30 cycles) without generating any side bands.

Extreme High Sensitivity
< Example 1: Amplification in the F3 speed level >
The results of amplifying 469 bp GAPDH from human genomic DNA in the F3 speed level are shown in this example. The PCR reaction volume was 20 μL and amount of the template DNA was varied from 10 ng down to 0.03 ng, corresponding to from 3,000 copies to 10 copies. PalmTaq™ High-Speed DNA polymerase (Part No. RK1010 and RK1020) was used. Speed of the thermal cycling in the F3 speed level is 48 seconds/cycle, corresponding to 24 minutes/30 cycles.

After the 30-cycle run in the F3 level as shown, the target amplicon bands are observed with strong intensity for all template concentrations. Down to 300 copies, the amplified band shows near saturated level of band intensity. Even in the case of 30 copies of the template, the amplified band stays with significantly strong intensity, suggesting that the sensitivity can go down further.

< Example 2: Amplification in the T2 speed level >
In this example, 200 bp β-globin was amplified from human genomic DNA in the fastest speed level T2. The PCR reaction volume was 20 μL and amount of the template DNA was varied from 10 ng down to 0.03 ng, corresponding to from 3,000 copies to 10 copies. PalmTaq™ High-Speed DNA polymerase (Part No. RK1010 and RK1020) was used. Speed of the thermal cycling in the T2 speed level is 36 seconds/cycle, corresponding to 18 minutes/30 cycles.

After the 30-cycle run in the T2 level as shown, the PCR amplification is completed successfully for all template concentrations, generating target amplicon bands with sufficiently strong intensity. This example demonstrates that the Palm PCR system can deliver PCR amplification with ultra-fast speed and extreme high sensitivity down below 10 copies.

Wide Dynamic Range
< Example 1: Amplification in the F1 speed level >
This example demonstrates dynamic range of the PCR amplification in the F1 speed level. Target amplicons with size 204 bp – 1,563 bp were amplified from plasmid templates. The PCR reaction volume was 20 μL and amount of the template DNA was 10 pg. PalmTaq™ High-Speed DNA polymerase (Part No. RK1010 and RK1020) was used. Speed of the thermal cycling in the F1 speed level is 1 minutes/cycle, corresponding to 30 minutes/30 cycles.

After the 30-cycle run in the F1 level as shown, the target amplicon bands are observed with strong intensity up to about 1 kbp, and weak bands are observed above 1 kbp. Despite the very fast amplification in 30 minutes, the Palm PCR system provides dynamic range that is sufficiently broad for various applications of genetic identification purposes.

< Example 2: Amplification in the S1 speed level >
In this example, dynamic range of the PCR amplification in the slowest speed level S1 is demonstrated. Target amplicons with size 204 bp - 2,062 bp were amplified from plasmid templates. The PCR reaction volume was 20 μL and amount of the template DNA was 10 pg. PalmTaq™ High-Speed DNA polymerase (Part No. RK1010 and RK1020) was used. Speed of the thermal cycling in the S1 speed level is 1.3 minutes/cycle, corresponding to 39 minutes/30 cycles.

After the 30-cycle run in the S1 level as shown, the target amplicon bands are observed with strong intensity up to 2 kbp. Despite the fast amplification in 39 minutes, the Palm PCR system provides dynamic range that is sufficiently broad for most of the PCR applications.