Maximizing PCR and RT-PCR Success - Automatibility of PCR

PCR Brochure 10/2016 9 PCR additives Various PCR additives or enhancers are available for improving PCR results. It is claimed that these reagents relieve secondary DNA structure (e.g., in GC-rich regions or in long amplification products), lower template melting temperature, enhance enzyme processivity, stabilize DNA polymerases, or prevent attachment of polymerases to plasticware. Commonly used PCR additives include dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), and glycerol. Most QIAGEN PCR kits include the novel reagent Q-Solution® (see selection guides, pages 23–25), which changes the dynamics of the PCR reaction to increase sensitivity when amplifying GC-rich DNA and targets with strong secondary structures (Figure 10). M With Q-Solution Without Q-Solution – 4.8 kb Figure 10. Influence of Q-Solution on PCR success. A 4.8 kb fragment was amplified in standard reactions using TopTaq DNA Polymerase with or without Q-Solution. M: marker. Specific amplification was achieved only in reactions containing Q-Solution. Automatibility of PCR Reproducibility and standardization in each step of a PCR experiment is the basis for successful results. Reliable tools for automated setup of reactions and sensitive platforms for fragment detection are required to achieve this. PCR setup Manual reaction setup is error prone due to pipetting variability, which can result from incorrect pipet calibration and/or human error. This leads to inconsistencies between different experiments and labs. Maintaining pipetting precision and ensuring reproducibility is even more difficult when pipetting into 96- or 384-well plates. The results obtained can vary from lab to lab and from researcher to researcher. Comparison of results and subsequent downstream analysis is therefore challenging. Manual pipetting of PCR reagents increases the risk of contamination with nucleases – and this is especially critical when using RNA as a template. Manual pipetting is also time consuming and tedious, and can result in repetitive strain injury. QIAGEN’s automated platform for high- precision PCR setup To eliminate pipetting errors and to ensure consistency, speed, and reliability in your research, QIAGEN offers the QIAgility® – an automated platform for rapid, high- precision PCR setup in all formats. The innovative QIAgility can set up multiple master mixes and process multiple PCR setups in parallel, streamlining your research and increasing productivity. The risk of contamination is greatly reduced, errors associated with manual handling are minimized, and improved yields are achieved.

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