NUCLEIC ACIDS

OVERVIEW​ BENEFITS

• Purify >10 µg of low endotoxin plasmid DNA (~0.1 EU/µg) with concentrations >100 ng/µL
• Fully automated process from crude to desalted plasmids in under 60 minutes
• ​Templated workflows that combine automated ion exchange and size exclusion chromatography
• ​Delivered with pre-programmed software scripts with an intuitive user interface to control sample volumes, aspirate/dispense cycles and wash steps

Purifying nucleic acids from a sample is a crucial step in numerous molecular biology applications, and hands-free automation with IMCStips will help ensure efficiency and reliability using your automated liquid handler of choice.

IMCStips are pipette tip-based micro-chromatography columns that utilize patented dispersive solid-phase extraction (dSPE) technology to provide transfection-grade plasmid DNA purification for your miniaturized samples ("mini preps" or "minipreps"). Our patented dSPE technology leverages silica or strong cation exchange resins, making it possible to bind more of your analytes in less time. By combining this step with buffer exchange through size exclusion chromatography, IMCStips and our automation expertise provide a one-stop shop for nucleic acid purification.

MAM WORKFLOW USING SIZEX IMCSTIPS^®

Automated Plasmid Purification with
µPure IMCStips​

µPure IMCStips® uses dispersive solid-phase extraction (dSPE) technology, offering a reliable and automated solution, capable of hands-free processing up to 96 samples in less than 55 minutes. Ensure consistent and dependable results with yields over 10 μg and purity that matches traditional methodologies, all while maintaining higher recoveries and purity compared to conventional magnetic bead kits.

Comparative Analysis of Plasmid DNA Purification Methods Across Different Plasmid Sizes. A) Total yield of plasmid DNA (μg) isolated using three different purification methods: Spin plate, Magbeads, and IMCStips. B) Purity of plasmid DNA as indicated by the 260/230 nm absorbance ratios, with the “expected” optimal purity threshold between 2.0 and 2.3 highlighted by the dashed lines. C)  Absorbance ratio at 260/280 nm, a further indicator of DNA purity, with points representing individual samples and the dashed lines marking the optimal range (1.8-2.0) also was impacted by purification kit. Data points are color-coded by the plasmid sizes (● 4k, ● 5k, ● 8k) used in the transformation, and the error bars reflect the standard deviation, underscoring the reproducibility of each purification method.

Spin plates and IMCStips® yielded similar purities and yields across various plasmid sizes, while magbead-based purifications were inaccurately high as reported by NanoDrop, likely due to contaminants. Absorbance ratios at 260/280 nm and 260/230 nm for magbead purifications did not fall within the expected range, indicating potential purity issues. In contrast, gel analysis confirmed that spin plates and IMCStips® provided consistent yield, purity, and supercoiled DNA content.

Comparison of tip-purified (1 mL IMCStips® packed with silica resin) to spin plate- and mag bead-purified pDNA. Figures A and B show 1 µL of eluted pDNA for the IMCStips, Spin Plate, and Magbeads methods for two plasmid sizes (4k, and 8k). Figures C and D show normalized gel electrophoresis for direct comparison of purification methods, adjusting volumes to 3.75 µL for IMCStips and Spin Plate and maintaining 1 µL for Magbeads to equalize pDNA concentrations.

Automated Low-Endotoxin Plasmid Purification 

with µPure LE IMCStips®

µPure LE IMCStips® utilizes patented dispersive solid-phase extraction (dSPE) technology for the high-throughput sample preparation of low-endotoxin, transfection-grade plasmid DNA. The tips are packed with loose resin that is dispersed within the sample, leading to turbulent mixing and improved binding. This automated workflow enables you to purify up to 96 samples in less than one hour.

Plasmid DNA were purified from overnight cultures of plasmid+ LB media using microPure LE IMCStips (tip size: 1 mL) followed by buffer exchange into 1x TE Buffer with SizeX100 IMCStips (P/N: 04T-H6R76-0A-220-8 or -96). Average amount of plasmid amounted to 14.6±1.3 μg, 10.9±1.1 μg, and 11.3±1.1 μg of plasmid for pCRS158, pCRS166, and pCRS240.3 respectively (A, and Table). Average concentration for all three plasmids were greater than 100 ng/μL (B), while volumes of eluate ranged from 90 μL to 112 μL after buffer exchange with SizeX100 (C). Agarose gel showing purified plasmid DNA from pCRS158 (blue), pCRS166 (red), and pCRS240.3 (green) without BamHI (D) and with BamHI (E).

Automated PCR Cleanup with IMCStips®

IMCStips for PCR cleanup is based on the traditional selective binding of double-stranded DNA to silica-based resin in the presence of chaotropic salts, with elution in a low salt buffer. IMCStips rapidly isolate target analytes within the pipette tips by leveraging the dispersive solid phase extraction approach with automated liquid handlers during aspiration and dispense steps. This approach simplifies automation by eliminating the need for ancillary equipment such as vacuum manifolds, centrifuges, or magnetic plates. Process up to 96 samples in 30 minutes or less.

Sanger sequencing results for one of the PCR products (608 bp). All five purified products were sequencing compatible.