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High Throughput Plasmid DNA Preparation Using A Magnetic Bead Kit On An Automated Workstation

arting Materials:

- DH10B Cells
- 1 Rack of 100 μL tips per sample plate
- 1 Rack of 200 μL tips per sample plate

Sciclone Procedure

Methodology

1. Attach 100 μL tips
2 . Add 95 μL Resuspension Solution and shake @ 1200-1500 rpm for 4 minutes
3. Add 100 μL Lysis Solution and shake @ 800-1000 rpm for 5 minutes
4. Add 100 μL Neutralization Solution and shake @ 800-1000 rpm for 10 minutes
5. Slowly dispense 150 μL of air to gently bubble material away from the tips. This is accomplished by lowering the tips to a height just below the top of the well, then moving back to the 12 o'clock position and lowering the tips to the bottom of the well and dispensing the air
6. After a 3 second pause, aspirate 110 μL of Lysate and transfer to a 96-well round bottom microplate
7. Replace 100 μL tips with 200 μL tips
8. Transfer 80 μL Isopropyl Alcohol, 10 μL air gap, and 10 μL Microparticle Solution to the Lysate plate
9. Mix the solution using 10 aspirate/dispense cycles
10.Place the round bottom plate on the magnet plate and allow the microparticles to settle for 10 minutes
11. Transfer all of the solution to waste taking care to not disturb the ring of microparticles
12.Add 170 μL 70% Ethanol allowing to rest for 30 seconds and then transfer to waste
13.Repeat step 12 two more times

Results

The plasmids prepared on the Sciclone were compared with plasmids prepared on Agencourt's DNATrack System. Table 1 shows the sequencing data for each system. The devices compare very closely to each other, indicating the Sciclone procedure is p
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