Maximizing PCR and RT-PCR Success

10 PCR Brochure 10/2016 Analysis of PCR fragments Post-PCR analysis and detection is commonly performed using agarose gel electrophoresis. Traditional agarose gel electrophoresis is time consuming and labor intensive, especially if there are large number of samples to be analyzed. Gel preparation also involves exposure to hazardous chemicals such as ethidium bromide. Thorough analysis of data in terms of fragment sizes and concentration is challenging, especially when data are to be compared with previously analyzed PCR products. Several factors such as the agarose quality and the percentage of agarose used affect the duration of electrophoresis and can influence results. Use of a high voltage during an electrophoretic run often results in smearing of nucleic acids, making analysis of results difficult. Standardization is of key importance when comparing data from different gel runs and this places a great emphasis on accurate electrophoresis conditions and record keeping. QIAGEN’s automated platform for effortless DNA fragment analysis For automated, high-resolution capillary electrophoresis, QIAGEN offers the QIAxcel Advanced System. DNA fragment analysis of 12 samples can be performed in as little as 3 minutes (Figure 11). Ready-to-run gel cartridges allow 96 samples to be analyzed with a minimum of hands-on interaction, reducing manual handling errors and eliminating the need for tedious gel preparation. With a resolution of 3–5 bp for fragments smaller than 0.5 kb, the QIAxcel Advanced System ensures greater accuracy than slab-gel methods, as well as greater confidence in data interpretation. Hands-free sample loading and self- contained components minimize exposure to hazardous chemicals such as ethidium bromide. Figure 11. QIAxcel Advanced process. Nucleic acid molecules are size separated by applying a current to a gel-filled capillary, and detected as they migrate toward the positively charged terminus. The signal data pass through a photomultiplier and are converted to an electropherogram and gel image by the QIAxcel ScreenGel Software. Gel matrix with dye Positive charge Capillary Nucleic acid with dye LED light source Detector Photo- multiplier QIAxcel ScreenGel Software Negative charge

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