Suitable for multiplex amplification, compatible with templates of varying GC% content, with good amplification uniformity and a wide range of blood input volumes.
Six-plex direct amplification using the Qihengxing Blood Direct Amplification Kit and the Supplier T kit was performed using fresh EDTA-anticoagulated blood at different concentrations, including 5%, 10%, 15%, 20%, 25%, and 30%. The amplified fragments ranged from 200 to 800 bp, with a GC percentage range of 35% to 77%. Amplification was performed for 35 cycles, and PCR products were analyzed by agarose gel electrophoresis. The results showed that the Qihengxing Blood Direct Amplification Kit successfully amplified all six primer pairs with high product yields and good amplification uniformity, using fresh EDTA-anticoagulated blood at different concentrations, ranging from 5% to 30%. However, the Supplier T kit failed to amplify all six bands, exhibiting low product yields and poor uniformity. The Qihengxing Blood Direct Amplification Kit exhibits greater tolerance to inhibitors and outperforms the Supplier T kit in amplification performance.
Compatible with different EDTA blood samples: fresh blood, 4℃, room temperature and -20℃ frozen blood samples
Fresh EDTA-anticoagulated blood was incubated at room temperature for 5 minutes, 1 day, 2 days, and 5 days. Six-plex direct PCR amplification was performed on each of these blood pairs using the Qihengxing Blood Direct Amplification Kit. Blood was added at 10% of the reaction volume. After 35 cycles of amplification, the samples were centrifuged at 4000 rpm for 2–3 minutes, and the supernatant was subjected to gel electrophoresis. The results in the figure below demonstrate that the kit accurately amplified all six products under different room temperature incubation times.
Fresh EDTA-anticoagulated blood was stored at 4°C for 1, 2, and 5 days. Six-plex direct PCR amplification was performed using the Qihengxing Blood Direct Amplification Kit. Blood was added at 10% of the reaction volume. After 35 cycles of amplification, the supernatant was centrifuged at 4000 rpm for 2–3 minutes and subjected to gel electrophoresis. As shown in Figure 3 below, the kit accurately amplified all six products under the different storage times at 4°C.
Fresh EDTA-anticoagulated blood was stored at -20°C for 1 day, 2 days, 5 days, 55 days, 8 months, and 20 months, respectively, and then subjected to direct PCR 6-plex amplification using the Qihengxing Blood Direct Amplification Kit. Blood was added at 10% of the reaction volume. After 35 cycles of amplification, the supernatant was centrifuged at 4000 rpm for 2-3 minutes, and subjected to gel electrophoresis. The results shown below demonstrate that the kit accurately amplified all six products under the different storage times at -20°C.
