MagVigen™ Plasma DNA Capture nanoparticles are ideal for DNA capturing from plasma samples. MagVigen™ Plasma DNA Capture nanoparticles feature efficient recovery of double-stranded and single-stranded DNA. DNA molecules are captured by MagVigen™ Plasma DNA Capture nanoparticles following a short incubation. The generated nanoparticle-oligo complex can be separated from the rest of the sample by a magnet. The retained genetic material can be eluted from the nanoparticles using an elution buffer.
MagVigen™ Plasma DNA Capture nanoparticles enable the purification of DNA from blood samples such as plasma and serum. The purified DNA products can be further analyzed by gel electrophoresis, PCR quantification and sequencing.
- MagVigen™ Plasma DNA Capture nanoparticles
- Lysis Buffer
- Proteinase K Buffer
- Proteinase K Powder
- Wash Buffer 1 Stock
- Elution Buffer
- SDS, 20%
- Magnetic Rack (NVIGEN Cat# A20006)
The MagVigen™ Plasma DNA Capture Kit (K61003) is capable of capturing small circulating cell-free DNA (>30bp) from plasma and serum.
- Prepare the plasma samples: if using frozen plasma, thaw under room temperature. Centrifuge plasma at 12000x g for 5 mins to remove any blood and cell debris.
- Prepare Proteinase K Solution: Add Proteinase K buffer to Proteinase K Powder, vortexing to mix.
- Prepare Wash Buffer 1: Calculate needed quantity according to Table 1. Add 750 ul Ethanol to every 250 ul of Wash Buffer 1 stock. Fresh made buffer performs the best.
- Check Table 1 on the total volume to decide types of centrifuge tubes (1.5ml, 2ml, 5ml, 15ml or 50ml) to use.
- Add proteinase K to a sample tube, add plasma to the tube. Add 20% SDS to the tube. Vortexing for 1 min to mix. Briefly spin down. Add Lysis Buffer to the tube. Vortexing for 1 min to mix.
- Add MagVigen™ Plasma DNA Capture nanoparticles. Add Isopropanol according to Table 1, vortexing to mix well. Incubate at 65°C for 40 min.
- Put the reaction tube on a magnetic rack to pellet the beads until the solution is clear (5-10 minutes depending on the strength of the magnet and the sample volume).
- Slowly remove the supernatant. Be careful not to take any beads, remove as much solution as possible. Tap the magnetic rack on a solid surface to allow residual solution to settle down to tube bottom, remove all supernatant.
- Take tube off magnet, add Wash Buffer 1. Mix by pipetting or vortexing. Transfer the solution to a 1.5 or 2 ml tube if needed. Briefly spin down. Pellet the beads on the magnetic rack until the solution is clear (about 3 minutes), remove the supernatant. Tap the magnetic rack on a solid surface to settle all supernatant residue to the bottom of the tube. Remove all supernatant.
- Add Wash Buffer 2 (80% Ethanol), mix by pipetting or vortexing. Pellet the beads on the magnetic rack (about 2-3 min) and remove all supernatant. Tap the magnetic rack on a solid surface to settle all supernatant residue to the bottom of the tube. Remove all supernatant.
- Add Wash Buffer 3 (80% Ethanol), mix by vortexing. Pellet the beads (about 2-3 min) and remove all supernatant.
- Briefly spin down, pellet the beads on the magnetic rack, and completely remove all residual of supernatant. Leave tubes on magnetic rack and air dry the pellet to evaporate all ethanol. This takes 2-3 minutes.
- Add desired amount of Elution Buffer to the beads, pipette up and down until all pellet has re-dispersed completely.
- Set the tube on a magnetic stand for 2-3 minutes.
- Collect supernatant from the bottom of the micro-centrifuge tube carefully without disturbing the pellet. The supernatant contains the extracted DNA. To ensure clean DNA sample without magnetic particles, put the vial by the magnet when pipetting the DNA solution out for downstream experiments.
Table 1. Reagent volume used in each step.
|Wash Buffer 1||100||300||500||2||4|
|Wash Buffer 2 80% ETOH||150||450||750||1.8||1.8|
|Wash Buffer 3 85% ETOH||150||450||750||1.8||1.8|
|Total volume at Lyse/Binding||0.6ml||1.9ml||3.2ml||12.8ml||25.6ml|