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.

Features and
Benefits

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.

 

System
Configuration

MICROLAB STARlet Autoloading liquid handling workstation with eight (8) independent pipetting channels, one (1) 96 channel Multi-Probe Head and iSWAP labware manipulating arm
 Hamilton
173000-036
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
Instrument Controller 6436-01
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

Downloads

Application note

Links

 
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