Note: The product sheet online is for general guidance of how to use our products. The most updated product usage protocol will be included in each product ship-out.
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
- Proteinase K buffer
- 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 Lysis Buffer: Heat at 65°C for a few minutesif there is solid to make clear solution.
- Prepare Proteinase Solution: Add Proteinase K buffer to Proteinase K powder vial. Vortexing to mix well. Store at -20°C.
- Prepare Wash Buffer 1: Calculate needed quantity according to Table 1. Add 450 ul Isopropanol to every 550 ul of Wash Buffer 1 stock. Fresh make the buffer each time.
- 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 solution to the bottom of a sample tube, add plasma to the tube. Vortexing for 20s to mix well, briefly spin down.
- Add 20% SDS to the tube. Vortexing for 1 min to mix well, briefly spin down.
- Add Lysis Buffer to the tube. Vortexing for 1 min to mix well. Briefly spin down. Incubate at 60°C for 30 min.
- Add MagVigen™ Plasma DNA Capture nanoparticles, slightly shake the vial to disperse. Add Isopropanol, vortexing to mix well, briefly spin down, and keep vortexing for 60 min.
- Put the reaction tube on a magnetic rack to pellet the beads until the solution is clear (10-20 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(~ 10 minutes), remove the supernatant. Tap the magnetic rack on a surface. Remove all supernatant.
- Add Elution Buffer, disperse the beads pellet by pipetting or vortexing. Wait for 3 min. Pellet the beads on a magnetic rack and transfer the supernatant to a new 1.5 ml tube.
- Add Wash Capture Beads, mixing and incubate for 15 min, then pellet the beads on a magnetic rack (~ 5 min), remove the supernatant.
- Wash the beads pellet using 75% Ethanol, disperse the bead pellet by pipetting or vortexing. Pellet the beads on a magnetic rack (~3 min) and remove all supernatant. Tap the magnetic rack on a solid surface to settle supernatant residue to the bottom of the tube. Remove all supernatant.
- Wash the beads pellet again using 75% Ethanol, mix by pipetting or vortexing. Pellet the beads (~ 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 the 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. Keep beads dispersed in elution buffer for 3 minutes.
- Set the tube on a magnetic stand for 2-3 minutes.
- Collect supernatant from the bottom of the micro-centrifuge tube without disturbing the pellet. The supernatant contains extracted DNA. Put the tube by the magnet when pipetting the DNA solution out for downstream experiments. Avoid touching the tube bottom when pipetting the cfDNA extraction out for downstream assay.
Table 1. Reagent volume used in each step.
|Wash Buffer 1||100||162.5||325||650||975||1300|
|Wash Elution Buffer||8||12.5||25||50||75||100|
|Wash Cap. Beads||20||31||62||124||186||248|
|Wash Buffer 2 / 3 (75% Ethanol)||100||162.5||325||650||975||1300|
|Total volume at Lyse/Binding||0.6ml||1.4ml||2.8ml||5.6ml||8.4ml||11.2ml|