A Single Plate for Multiple Applications
Purified RNA or DNA is required for most molecular biology experiments and their isolation is often one of the first steps in disease pathways research. So RNA and DNA purification is a regular part of molecular biology and disease research.
As the throughput in research has increased dramatically, efficient purification of plasmid DNA, genome DNA and RNA isolation in multi-well plates has become a crucial need.
To address the needs of researchers working in these areas, Pall has recently launched the AcroPrep Advance 96-well Long Tip Filter Plate for Nucleic Acid Binding. One benefit of the new plate includes the incorporation of a silica-based quartz glass fiber media to allow for efficient binding of DNA and RNA, while providing smooth flow and rapid processing of samples. The benefit of adding this media is that it offers researchers the flexibility to purify plasmid DNA from bacteria, and genomic DNA or total RNA from cell culture samples, thus providing a single plate for multiple applications.
A unique aspect of the plate is the new outlet tip geometry that provides direct flow of samples into the receiver plate which minimizes hanging drop formation and reduces the possibility of cross contamination (Figure 1). Cross contamination causes inconsistent results and makes experiment reproducibility very difficult.
The plates are manufactured in accordance with specifications for the Society of Biomolecular Screening (ANSI/SBSx-2004) for multi-well plates allowing the entire DNA purification process to be performed on automated equipment. Thus enabling process automation, a key tool in disease research.
Specific Application Areas
Enables High Throughput Plasmid DNA Purification
With applications increasing in drug delivery, pre-clinical studies, and DNA vaccines, demand for high purity plasmid DNA has been on the rise. Furthermore, advances in cloning techniques have caused the number of samples to also increase. High sample numbers require the use of high-throughput small-scale plasmid preparation.
In a recent application note, Pall describes how the special features of their AcroPrep Advance 96-well Long Tip Filter Plate for Nucleic Acid Binding (Pall NAB plate) translate into higher throughput, increased sensitivity and signal-to-noise ratios, and reduced processing times in plasmid DNA purification.
The quartz membrane in the plate provides fast flow rates, efficient purification of target DNA, and removal of contaminants without clogging. The data showed that the Pall NAB plates allowed purification of plasmid DNA with good yield and high quality using generic laboratory reagents prepared in-house.
High quality of the purified plasmid DNA was demonstrated by evaluation of PCR amplification (Figure 2), restriction enzyme digestion (Figure 3), and Sanger sequencing (Figure 4).
DNA fragments of expected size were obtained following the PCR amplification and restriction enzyme digestion, and high quality sequencing results with long read lengths were obtained in Sanger sequencing, all confirming that the purified plasmid DNA was well suited for these downstream applications. The results are comparable with those obtained with plasmid DNA purified using commercially available kits, which indicates that the Pall NAB plates are able to generate plasmid DNA of the same quality as can be obtained with commercially available kits, but at greatly reduced cost.
For more information on other applications for the AcroPrep Advance 96-well Long Tip Filter Plate for Nucleic Acid Binding , please see the following application notes: