Qx dna size marker

A series of templates composed of different ratios of BN and LEW DNA, including 0%, 0.1%, 0.2%, 0.4%, 0.8%, 1%, 2%, 4%, 8%, 10%, 20%, 40%, 60%, 80%, and 100% BN DNA, were prepared as templates for D10Rat25-PCR. Following PCR amplification, capillary electrophoresis was performed to resolve the products, whose sizes were determined by referencing the signals of a QX Alignment Marker and QX DNA Size Marker (QIAGEN, Valencia, CA). For each template, signal intensities stemming from BN and LEW D10Rat25 were obtained, and the ratio of BN/(BN+LEW) was derived. The real percentage of DNA was plotted against the corresponding signal ratio, and a standard curve was derived by linear regression with a correlation coefficient (R²).

PCR products were sized using QIAxcel DNA Screening gel cartridge (QIAGEN) on the QIAxcel system (QIAGEN, Switzerland), which enabled high-resolution capillary electrophoresis. A 50–800 bp QX DNA Size Marker (Qiagen) was included on QIAxcel runs, and the size of the products was determined using the QIAxcel ScreenGel software (Qiagen). The QIAxcel system produces a digital gel image and an electropherogram for fragment analysis. The expected PCR product sizes of set A were 749bp (HMPV), 534 bp (HADV), 375 bp (HCOV-229E/NL63), 264 bp (HPIV-2), 188 bp HPIV-3), and 139 bp (HPIV-1), while the product sizes of set B were 754 bp (IFV-B), 578 bp (HCOV-OC43/HKU1), 394 bp (HRV-A/B), 273 bp (HRSV-A), 206 bp (IFV-A) and 143 bp (HRSV-B).