Fully-Automated Method for Purifying PCR* Products using Promega's Wizard
MagneSil* PCR Clean-Up System on the Biomek® FX
Liquid Handling System
**Chad Pittman, ***Graham Threadgill, Ph.D., and **Laura Pajak, Ph.D.
Beckman Coulter, Inc.
**Indianapolis, Indiana and ***Fullerton, California
Brad Larson and Paul Otto
Promega Corporation - www.promega.com
Madison, Wisconsin
| Q. | How many plates are processed using this method? |
| A. | Two plates are processed on the worksurface of the Biomek FX in approximately one hour. |
| Q. | What Biomek FX configuration is required for this method? |
| A. | The method will run on a Biomek FX that has a left pod with a 96-multichannel p200 head. The method was developed on a hybrid system (multichannel/Span 8) to allow for Quantitation and Normalization however the Span 8 pod is not used during the purification process with MagneSil reagents. Additionally, the following ALPS: a trash ALP, a MagBead ALP, an Orbital Shaker ALP and a 96 Tip Wash Station, are utilized during processing. |
| Q. | Is there a time limit on how long the beads can sit at room temperature? |
| A. | The beads are stable at room temperature if covered to minimize evaporation. However, for long term storage, it is recommended that the particles be placed back into the supplied bottle and freshly decanted for the next purification procedure to ensure that the particles are efficiently resuspended during the method. |
| Q. | Do I have to do all the recommended washes? |
| A. | For complete contaminant removal it is recommended that all the ethanol wa |
| Q. | What is the benefit of the second capture and can I remove it? |
| A. | The second capture is used to remove as much bead carryover as possible. If particle carryover is not a concern, then the second capture may be removed to shorten the method time. |
| Q. | Can the Reagent-to-PCR product volume ratios be changed? |
| A. | It is recommended that the ratios in the method stay the same for optimal results. |
| Q. | Can the shake times be decreased throughout the method? |
| A. | Shaking the samples for the given time allows for optimal binding and removal of impurities. Decreasing the time may result in diminished product or inadequate results. |
* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffman-LaRoche, Ltd.
©1998 - 2006 Beckman Coulter, Inc.
