Genomics

DNA Purification Methods

Binding to Microporous Matrices

Examples of this method are:

• The use of high salt concentrations in the suspension buffer to bind DNA to silica matrices followed by elution using low salt concentrations in the elution buffer.
• Ion exchange chromatography using anion exchange membranes.

Pall has developed unique membrane technology in the form of Mustang™ anionic and cationic membrane adsorbers, which allows purification of nucleic acids and proteins using ion exchange chromatography. The Mustang product line is already established as the technology of choice for nucleic acid removal from process streams, as in part of DNA clearance from therapeutic products like vaccines. The utility of Mustang membrane adsorber modules has also been demonstrated for purification of plasmid DNA, for use as a gene therapy product. Another novel product developed by Pall to streamline nucleic acid purification is the plasmid lysate clearance plate which allows purification of plasmid DNA. For example, these plates can be used as a vector for archiving eukaryotic sequences as part of a bacterial or yeast clone library.

Size Exclusion

Ultrafiltration (UF) is a membrane separation technique used to separate extremely small particles and dissolved molecules in fluids. The primary basis for separation is molecular size, although other factors such as molecule shape and charge can also play a role. Compared to non-membrane processes, ultrafiltration is gentle, fast, and relatively inexpensive. Methods for purifying nucleic acids using the Pall Nanosep® device, which contains a membrane operating by molecular weight size exclusion, are well established and covered in the application guide, “Nanosep Centrifugal Devices Protocols for Use.” The methods described allow rapid, simple recovery of high yields of purified DNA. The same technology is also available in multi-well plate formats for parallel processing of biological samples requiring purification.

Chemical Extraction

These methods use solvents such as chloroform/phenol to solubilize cellular material and partition the DNA for subsequent purification, e.g., using density gradient centrifugation. The limitation of this method is the disposal issue created by the extraction solvents used and the purity and yield of the DNA recovered.

Sample Prep Products for DNA Sequencing

Remove Cell Debris to Enrich for Plasmid DNA

• AcroPrep™ Filter Plates for Lysate Clearing
• AcroPrep Filter Plates for Desalting
• Nanosep® Centrifugal Devices for Desalting

Sequencing Clean-up to Remove Unincorporated Label

Use hydrophilic microfiltration (e.g., Supor® or GHP membrane) filter plates or spin devices with resins to capture purified sequencing product.

• AcroPrep Filter Plates for Sequencing Clean-up
• Nanosep Centrifugal Devices for Sequencing Clean-up

PCR Clean-up to Remove Debris

Use ultrafiltration size exclusion plates or spin devices to directly remove debris leaving purified product in the filter well/tube. Use hydrophilic microfiltration (e.g., Supor or GHP membrane) filter plates or spin devices with resins to capture purified PCR product.

• AcroPrep Filter Plates for PCR Clean-up
• Nanosep Centrifugal Devices for PCR Clean-up

Sample Preparation in Genomics

Pall manufactures membranes for the purification and concentration of biomolecules that exhibit extremely high yields with low biomolecule binding. We incorporate these membranes into devices that use fast, gentle methods and minimize handling to protect your valuable samples. Our attention to product quality and performance is expressed in reproducible results and pure samples for downstream processing.

For concentration and purification steps requiring ultrafiltration, we offer Omega™ modified polyethersulfone membrane. This membrane is well documented for low non-specific protein and DNA binding, high recoveries, and superior chemical and physical properties.

For applications such as lysate cleaning, drug binding studies and bead-based assays, choose Bio-Inert® modified Nylon 6,6, GHP hydrophilic polypropylene, or Supor® modified polyethersulfone membrane, all of which are formulated for low binding and low extractables.

Pall has developed unique membrane technology, in the form of Mustang™ anionic and cationic membrane adsorbers, which allows purification of nucleic acids and proteins using ion exchange chromatography. The Mustang product line is already established as the technology of choice for nucleic acid removal from process streams, such as part of DNA clearance from therapeutic products like vaccines. The utility of Mustang membrane adsorber modules has also been demonstrated for purification of plasmid DNA, for use as a gene therapy product.

For single sample or low throughput applications, Pall's centrifugal devices provide simple, fast concentration of samples from 50 μL to 60 mL, with recoveries typically greater than 90%. For high throughput applications, choose Pall filter plates. Each of our filter plate platforms targets specific application challenges.

Detection in Genomics

Detection techniques are important for interpreting results in all areas of genomics research and molecular diagnostic applications. As sample sizes become smaller and more valuable, the need intensifies for highly sensitive detection platforms that exhibit extremely low background. Our inspiring portfolio of membranes and materials provides the superior sensitivity you need to detect minute genomic samples. These products are proven and trusted for gene expression analysis.

Pall manufactures all of our membranes to the strictest quality standards, assuring consistent performance from lot to lot and blot to blot. Biodyne® nylon membranes are available with a neutral charge, positive charge, or negative charge to meet the needs of various techniques. These membranes can be efficiently stripped and reprobed, and perform optimally with all detection systems. BioTrace™ NT is a pure, unsupported nitrocellulose membrane that is strong and durable, and features high nucleic acid binding capacities.

For increased throughput, choose our filter plates for various biomolecule-binding assays.

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