It is suitable for multiple amplification, compatible with templates with different GC% contents, has good amplification uniformity, and has a wide range of blood input amounts.
The Qihengxing Blood Direct Amplification Kit and the Supplier T Kit were used to perform direct amplification of 5%, 10%, 15%, 20%, 25%, and 30% fresh EDTA anticoagulated blood with 6 primers, respectively. The amplified fragment range was 200-800bp, the GC% range was 35%-77%, and 35 cycles were amplified. The PCR products were taken for agarose gel electrophoresis detection. The results showed that the Qihengxing Blood Direct Amplification Kit was able to successfully amplify the multiple amplification of 6 pairs of primers with different proportions of fresh EDTA anticoagulated blood input from 5% to 30%, with high product quantity and good amplification uniformity; while the Supplier T Kit failed to amplify all six items, and the product quantity was low and the uniformity was poor. The Qihengxing Blood Direct Amplification Kit has stronger tolerance to inhibitors and better amplification performance than the Supplier T product.
Compatible with different EDTA blood samples: fresh blood, 4℃, room temperature and -20℃ frozen blood samples
Take fresh EDTA anticoagulated blood and place it at room temperature for 5 min, 1 d, 2 d and 5 d, respectively. Use Qihengxing Blood Direct Amplification Kit to perform direct PCR 6-plex amplification on these blood sample pairs. The amount of blood input is 10% of the reaction volume. After 35 cycles of amplification, centrifuge at 4000 rpm for 2-3 min and take the supernatant for gel electrophoresis. The results in the figure below show that under different time conditions at room temperature, this kit can accurately amplify the products of all six items.
Fresh EDTA anticoagulated blood was taken and placed at 4℃ for 1 d, 2 d, and 5 d. The blood samples were directly amplified by 6-plex PCR using the Qihengxing Blood Direct Amplification Kit. The amount of blood input was 10% of the reaction volume. After 35 cycles of amplification, the supernatant was centrifuged at 4000 rpm for 2-3 min and gel electrophoresis was performed. As shown in Figure 3 below, under the conditions of different placement times at 4℃, this kit can accurately amplify the products of all six items.
Fresh EDTA anticoagulated blood was taken and placed at -20℃ for 1 day, 2 days, 5 days, 55 days, 8 months, and 20 months. The blood samples were directly amplified by 6-plex PCR using the Qihengxing Blood Direct Amplification Kit. The amount of blood input was 10% of the reaction volume. After 35 cycles of amplification, the supernatant was centrifuged at 4000 rpm for 2-3 minutes and gel electrophoresis was performed. The results in the figure below show that under the conditions of different storage times at -20℃, this kit can accurately amplify the products of all six items.
