SV 96 Plasmid DNA Purification
Plasmid DNA is useful in a wide range of molecular biology applications. However, purification of plasmid DNA from pelleted bacterial culture can be time-consuming when done manually. Automation of this process can deliver significant increases in throughput as well as higher levels of precision and accuracy. We have developed an automated method that enables processing of multiple 96-well plates per day using the Promega Wizard SV 96 Plasmid Purification System on the Hamilton MICROLAB STAR workstation. Here we provide an example of high-copy plasmid DNA purification with this easy-to-follow automated protocol. High-purity plasmid DNA is isolated consistently from bacterial culture samples with no detectable cross-contamination, and can be used directly in downstream sequencing and restriction enzyme digestion.
High-copy plasmid DNA was purified using the automated Wizard SV 96 Plasmid DNA Purification System method on the MICROLAB STAR workstation. Purity and yield were assessed via absorbance measurements using a Nanodrop* ND-1000 Spectrophotometer (Thermo Fisher, USA). Average yield of high-copy plasmid DNA (phMGFP) from MDS42 E. coli bacteria, based on 24 samples taken at an OD600 of 4, was 8.44 ± 0.93µg with an average concentration of 105.5 ± 11.6 ng/µl. Average purity of the isolated high-copy plasmid, based on 24 samples A260/280 and A260/230 measurements, was 1.92 ± 0.04 and 2.35 ± 0.20, respectively. For cross-contamination evaluation, the average yield of DNA from 8 blank well samples was also measured. Blank sample purity was found to be negligible at 0.01 ± 0.05µg.
|MICROLAB STARlet Autoloading liquid handling workstation with eight (8) independent pipetting channels, one (1) 96 channel Multi-Probe Head and iSWAP labware manipulating arm
|DWP Carrier (5 SBS positions)||182090|
|MTP Carrier (5 SBS positions)||182365|
|Disposable Tip Carrier (5 racks, 480 tips)||182085|
|Basic Vacuum System (BVS) include vacuum pump, controller, and collection bottle)||190033|
|Teleshake Shaker for DWP||187149|
|VENUS ONE Software||911004-09|
|300 µl CO-RE disposable filter tips in blue hanging rack||235903|
|1,000 µl CO-RE disposable filter tips in blue hanging rack||235905|
|96-well deep-well pyramid-bottom reservoir plates||56669-01|
|12-column deep-well reservoir plates||Promega V6791|
|Wizard® SV 96 Plasmid DNA Purification System||Promega A2250|