GeneRulor NGS DNA Purification Magnetic Beads

1.DNA Purification

(1) Take the magnetic bead solution out from 2~8 °C 30 minutes inadvance, and let it stand to equilibrate to room temperature.

(2) Invert or vortex the magnetic bead solution to mix it thoroughly. Pipette an appropriate volume of the bead solution into the DNA sample according to Table 1, and gently pipette up and down 10 times to mix well.

(3) Incubate at room temperature for 10 minutes to allow DNA to bind to the magnetic beads.

(4) Place the sample on a magnetic rack, and carefully remove the supernatant after the solution becomes clear (approximately 5 minutes).

(5) Keep the sample on the magnetic rack at all times, add 200 μL of freshly prepared 80% ethanol to rinse the magnetic beads, incubate at room temperature for 30 seconds, and carefully remove the supernatant.

(6) Repeat Step 5 once for a total of two rinses.

(7) Keep the sample on the magnetic rack at all times, and air-dry the magnetic beads with the tube cap open at room temperature for approximately 5-10 minutes.

(8) Remove the sample from the magnetic rack, add an appropriate volume of nuclease-free water, vortex or pipette up and down to mix thoroughly, and let it stand at room temperature for 2 minutes. Place it on the magnetic rack for 5 minutes until the solution is clear, then carefully transfer the supernatant to a new nuclease-free centrifuge tube.

2. Reference Conditions for DNA Purification

Figure 1. Recovery results of 100 bp marker with different magnetic bead dosages

3. DNA Fragment Size Selection

(1) Take the magnetic bead solution out from 2~8 °C 30 minutes inadvance, and let it stand to equilibrate to room temperature.

(2) Vortex the magnetic bead solution to mix it thoroughly. Pipette an appropriate volume of the bead solution (for the first round of selection, refer to Table 2) into the purified DNA sample, and gently pipette up and down 10 times to mix well.

(3) Incubate at room temperature for 10 minutes to allow DNA to bind to the magnetic beads.

(4) Place the sample on a magnetic rack, and carefully transfer the supernatant to a new nuclease-free centrifuge tube after the solution becomes clear (approximately 5 minutes).

Note: It is recommended to leave 2 μL of liquid at the bottom of the tube when transferring the supernatant to avoid aspirating magnetic beads and affecting the selection effect.

(5) Add an appropriate volume of magnetic bead solution (for the second round of selection, refer to Table 2), and pipette up and down 10 times to mix well.

(6) Incubate at room temperature for 10 minutes to allow DNA to bind to the magnetic beads.

(7) Place the sample on a magnetic rack, and carefully remove the supernatant after the solution becomes clear (approximately 5 minutes).

(8) Keep the sample on the magnetic rack at all times, add 200 μL of freshly prepared 80% ethanol to rinse the magnetic beads, incubate at room temperature for 30 seconds, and carefully remove the supernatant.

(9) Repeat Step 8 once for a total of two rinses.

(10) Keep the sample on the magnetic rack at all times, and air-drythe magnetic beads with the tube cap open at room temperature for approximately 5-10 minutes.

(11) Remove the sample from the magnetic rack, add an appropriate volume of nuclease-free water, vortex or pipette up and down to mix thoroughly, and let it stand at room temperature for 2 minutes. Place it on the magnetic rack for 5 minutes until the solution is clear, then carefully transfer the supernatant to a new nuclease-free centrifuge tube.

4. DNA Fragment Selection Reference Conditions

Figure 2. Electropherogram on Agilent 2100 High Sensitivity DNA Chip. Smear fragments were dissolved in ddH₂O, and fragment size selection was performed with magnetic beads at volume ratios from 0.80×/0.20× to 0.45×/0.15×