Biotage Miniprep PhyTip® Columns for Plasmid DNA purification technology provide mini scale plasmid DNA purification in a 96 well format on our benchtop AutoPlasmid MEA and other liquid handling robots. The plasmid recovered requires no desalting or precipitation, yet they are transient transfection quality. This is accomplished using the PhyTip Dual Flow Chromatography technology with a proprietary silica resin. Unlike anion exchange, which requires desalting or precipitation, the plasmid recovered from the AutoPlasmid MEA has low endotoxin and may be used directly for any application including transient transfection. The process is a robust process, capable of performing regardless of sample variation (which is essential for automation).
The development of columns and automated plasmid miniprep method, for use as a benchtop system, has remained a challenge in the automation industry. The complex lysis and neutralization steps involved in almost every plasmid purification require skilled laboratory professionals to perform the sensitive procedures. In addition to the long time required to perform typical plasmid purifications, problems such as clogging, low yields, and carry over contamination can readily occur if care is not taken. The automated lysis and neutralization steps have been fully optimized starting from the cell pellet, resulting in a sensitive, robust method yielding samples suitable for transformation, transfection, and sequencing.
MiniPrep on the AutoPlasmid MEA
The AutoPlasmid MEA benchtop instrument may purify up to 48 samples per run, processing the samples 12-at-a-time. Mini prep scale plasmids are prepared directly from the cell pellet with a bench top instrument.
|STARTING CULTURE VOL. (ML)||MEDIUM||YIELD (µg)||AUTOPLASMID MEA||96-CHANNEL LIQUID HANDLING AUTOMATION|
Miniprep columns are available in tip volume (mL) of 1 and resin volumes (µL) of 80|300 depending upon robot configuration.
The pace of innovation from life science researchers is constantly pushing the limits of available tools. While technologies such as high throughput construct screening, crystallography and DNA sequencing have advanced, techniques for performing plasmid DNA minipreps have remained fairly unchanged. At the same time, greater emphasis is being placed on utilizing automated liquid handling systems to adapt traditional plasmid DNA purification formats like filter plates and magnetic beads to higher throughput, but with limited success. The major challenge to these approaches lies in the complexity of the lysed sample. Current methods require removal of precipitates and debris from bacterial cell lysate prior to DNA binding step by centrifugation or vacuum filtration and “careful transfer.” Centrifugation is difficult to automate and requires human intervention. Filtration and transfer methods often result in clogged pipet tips, transfer of precipitates to filters, inconsistent fluid flow, and cross contamination between samples. Such obstacles to high throughput have made currently available methods for automating plasmid DNA minipreps less than satisfactory. PhyNexus has developed a solution for obtaining reproducible automated plasmid DNA minipreps in a truly walk-away fashion. Lysate Direct PhyTip® Column technology consists of a purification column in a pipette tip format. These novel purification columns are composed of pipette tip containing plasmid DNA binding resin and a thin, inert frit screen at the end of the tip to retain the resin. The unique design of the column allows for DNA binding directly from cell lysates in the presence of precipitates and debris without sample clearing. By eliminating the need for centrifugation or filtration, Lysate Direct PhyTip® Columns for Plasmid DNA Purification enable utilization of liquid handing automation found in high throughput laboratories to easily purify up to 96 samples in parallel. Lysate Direct PhyTip® Columns are also compatible with the MEA Personal Purification System available from PhyNexus for 12-at-a-time sample processing.
Robot Resin Part Number Description PhyNexus Autoplasmid MEA
Lysate Direct 80 µL PhyTip columns for plasmid DNA 1 box kit
PhyNexus Autoplasmid MEA
Lysate Direct 40 µL PhyTip columns for plasmid DNA 1 box kit
PhyNexus Autoplasmid MEA
Lysate Direct 80 µL PhyTip columns for plasmid DNA 10 box kit