Maximizing PCR and RT-PCR Success - Ease of use

12 PCR Brochure 10/2016 Ease of handling Many PCR reagents include a number of additional features (e.g., master mixes, visualization dyes, or convenient room-temperature setup) for streamlining the PCR procedure and reducing the risk of handling errors. QIAGEN offers a range of kits providing these features as well as dNTPs and integrated gel loading dye for further time savings (Figure 14). Hot-start PCR enzymes that exhibit no activity at room temperature are available for convenient PCR and RT-PCR setup. In addition, a novel enzyme-stabilizing additive in TopTaq DNA Polymerase Kits improves the ease of setup when compared with standard PCR enzymes and allows the enzyme to be stored in a refrigerator – eliminating the need to freeze and thaw the reagent. For a comprehensive overview of the exceptional handling features of QIAGEN PCR and RT-PCR products, see pages 23–25. Ease of use In addition to factors such as the choice of enzyme, reaction conditions, template quality and primer design, researchers must also consider of use and convenience during reaction setup. The robustness of a particular PCR reaction under various conditions – such as different salt concentrations, annealing temperatures, and cycle numbers – must be evaluated. Figure 13. Comparison of manual reaction setup with automated setup using QIAgility. QIAxcel readout of one-step RT-PCR–amplified 295 bp actin beta (ACTB) mRNA target. The QIAGEN OneStep Ahead RT-PCR Kit provides superior sensitivity and specificity compared with other suppliers. Manual versus automated setup with the QIAGEN kit yields comparable results, while kits from other suppliers yield poorer results with automated setup, due to the longer exposure to room temperature during the automated setup process. Manual QIAGEN OneStep Ahead RT-PCR Kit Supplier T Supplier P Supplier TI QIAgility 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC 10 ng 100 ng 1 pg NTC

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