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Application of MEDUSA 96-channel pipette in plasmid DNA extraction from magnetic beads

Application of MEDUSA 96-channel pipette in plasmid DNA extraction from magnetic beads
MagPure Plasmid Mini Kit Operating Procedure
Ready to work
● 96-channel pipetting gun MEDUSA96
● IKA MS3 96-well oscillator
● 96-hole deep hole plate
● 96-well plate magnetic stand
● 96-well plate centrifuge Hettich 320
● MAGEN MagPure Plasmid Mini Kit Plasmid Extraction Kit
Operating procedures
1. Incubate 1.2 ml of bacterial bacterial solution in a 96-well plate to the appropriate concentration according to standard procedures. (In this experiment, in order to verify the parallelism of this method for each sample, 250 ml of bacterial solution was used, and then 1 ml was transferred to each well with a 96-channel pipette)
2. Centrifuge the bacteria at 2,000 xg for 5 minutes and discard the medium.
3. Transfer 170 ul of Buffer P1/Rnase A to the bacterial pellet using a 96-channel pipette. Place on the shaker and shake vigorously at 1500-2000 rpm for 1-2 minutes.
4. Transfer 170 ul of Buffer P2 to the bacterial resuspension using a 96-channel pipette. Place on a shaker and mix at 300-600 rpm for 3 minutes to lyse the bacteria.
5. Transfer 170 ul Buffer N3 to the bacterial lysate using a 96-channel pipette. Place on a shaker and mix by shaking at 300-600 rpm for 5 minutes.
6. Place in the centrifuge. The impurities were removed by centrifugation at 4000-4700 g for 15 minutes.
7. Carefully remove the 96-well plate and place it in the platform of the 96-channel pipetting gun. Adjust the depth of the insertion of the transfer gun and draw 450 ul of the supernatant into the new 96-well reaction plate. (If there is more precipitation, only transfer 400ul)
8. Add 0.5 volumes of Bind Beads to the sample using a 96-channel pipette. Place on a shaker and mix at 1000-1200 rpm for 1 minute. Allow to stand at room temperature for 10 minutes (handle low copy number of plasmids at 2-8 °C overnight).
9. Place the magnetic beads on the magnetic stand for 5 minutes. Aspirate the waste liquid with a 96-channel pipette (take more than 800 ul).
10. Add 600 ul of 70% ethanol to each well using a 96-channel pipette. Place on a shaker and mix at 1000-1200 rpm for 1 minute.
11. Place the magnetic beads on the magnetic stand for 3 minutes. Aspirate the waste liquid with a 96-channel pipette (take more than 650 ul).
12. Add 200 ul of 70% ethanol to each well using a 96-channel pipette. Place on a shaker and mix at 1000-1200 rpm for 1 minute.
13. Place the magnetic beads on the magnetic stand for 3 minutes. Aspirate the waste liquid with a 96-channel pipette (take more than 250 ul).
14. Dry at room temperature for 20 minutes.
15. Add 50-100 ul of sterilized water (Elution Buffer) to each well using a 96-channel pipette and place on a shaker at 1200-2000 rpm for 2 minutes.
16. Place the magnetic beads on the magnetic stand for 3 minutes. Transfer the plasmid solution to a new 96-well plate using a 96-channel pipette.
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