高品質の循環無細胞DNA（cfc-DNA）を簡便に抽出可能
Product Features
Figure 1. Purification of cell-free circulating DNA from different plasma volumes. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit (Cat# 55500) was used to purify circulating DNA from 50 µL, 100 µL and 200 µL plasma prepared from blood collected on citrate as an anticoagulant in comparison to Competitor Q's kit. Two microlitres of the purified DNA was then used as the template in qPCR reactions to assess the relative amount of the purified housekeeping 5S rRNA gene. The relative amount of the 5S rRNA gene increases linearly with increasing the sample input volume. Norgen's kit showed the most consistent and the highest recovery of the housekeeping 5S rRNA gene as compared to the other isolation method.
Figure 2. Linearity of DNA purified from increasing plasma volumes using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit (Cat# 55500). Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit as used to purify circulating DNA from 50 µL, 100 µL and 200 µL plasma prepared from blood collected on citrate as an anticoagulant. Two microlitres of the purified DNA was then used as the template in qPCR reactions to assess the linearity of the purified housekeeping 5S rRNA gene from the different plasma volumes. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit was able to recover 96% of the 5S rRNA gene from 100 µL plasma relative to the amount that is present in 50 µL plasma. Moreover, 97% of the 5S rRNA gene was recovered from 200 µL plasma relative to the amount that is present in 100 µL plasma.
Figure 3. Determination of the amount of inhibition present in plasma cell-free circulating DNA samples when detecting the human 5S gene. DNA was isolated from 50 µL, 100 µL and 200 µL plasma using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit (Cat# 55500). Increasing volumes of the elution (2, 4 and 8 μL) were used in a 20 μL qPCR reaction to observe any decrease in Ct value. An increase in Ct values with increasing amount of template would be a clear indication of PCR inhibitors present in the sample. An increase in elution volume used as a template in the qPCR did not affect the Ct value generated from qPCR and infact the Ct values tend to decrease with increasing the PCR input volume indicating that DNA purified from plasma using Norgen's kit is free of the common inhibitors usually present in plasma.
| Kit Specifications | Micro Kit (Cat.55500) | Mini Kit (Cat. 55100) |
| Minimum Plasma/Serum Input | 10 μL | 200 μL |
| Maximum Plasma/Serum Input | 200 μL | 500 μL |
| Size of DNA Purified | All sizes | All sizes |
| Elution Volume | 25-50 μL | 50-100 μL |
| Time to Complete 10 Purifications | 15-20 minutes | 15-20 minutes |
| Average Yields | Variable dependingon specimen | Variable dependingon specimen |
| Kit | Input | Cat. # |
| Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit | 10 μL – 200 μL | 55500 (50 preps) |
| Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit | 200 μL – 500 μL | 55100 (50 preps) |
Related Products
| Kit | Input | Cat. # |
| Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit | 1 mL – 4 mL | 55600 (20 preps) |
| Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit | 5 mL – 10 mL | 55800 (10 preps) |