OVERVIEW
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.
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
RESINS & APPLICATIONS
Kits for Nucleic Acid Purification:
- µPure LE Kit for Hamilton Systems with SizeX (Cat#: 04K-H001)
- µPure LE Kit for Hamilton Systems (Cat#: 04K-H002)
- IMCStips PCR Clean up Kit for Hamilton Systems (Cat#: 04K-H003)
View and download our product sheets and flyers below:
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.
Three replicates of tip-purified PCR products of varying sizes (221-1346 bp).
| 221 | 377 ± 163 |
| 443 | 851 ± 113 |
| 608 | 747 ± 108 |
| 740 | 776 ± 135 |
| 1346 | 1183 ± 243 |
The table summarizes the amount recovered for each PCR product.
Sanger sequencing results for one of the PCR products (608 bp). All five purified products were sequencing compatible.
Automated Midi Prep Scale Plasmid Purification with MidiPure™ IMCStips®
MidiPure™ IMCStips® on the Dynamic Devices platform offers an efficient solution for automated, high-throughput plasmid DNA purification. Designed for larger volumes, it enables labs to convert 5 mL pipetting systems into mid-scale purification platforms. This upgrade from the 1 mL μPure IMCStips protocol allows labs to process up to 500 mL of culture, isolating nearly 2.4 mg of plasmid DNA per plate in less than two hours.
Yield and purity of pDNA across different cell densities.(n = 4). 39 ± 1 μg of pDNA was isolated from 20 OD₆₀₀ and 41 ± 4 μg of pDNA from an OD₆₀₀ of 30. The yield increased for 50 OD samples to 55 μg of pDNA, but similar yields were obtained from OD 80 and 100 samples at 66 and 68 μg of pDNA. This suggests that 100 mg silica MidiPure is preferable for samples up to 60 OD, whereas 150 and 250 mg silica MidiPure can be used for higher loads.
Plasmid yield and purity comparing MidiPure IMCStips and a midi spin column. One MidiPure IMCStips (150 mg of silica per tip) yielded 117 ± 3 μg of plasmid, while one spin column from Company A isolated 251 ± 9 μg. Since the resuspension, lysis, and neutralization volumes were double of the spin column (58 mL final volume for IMCStips versus 30.8 mL final volume for spin column), the yield is comparable with IMCStips at approximately half the amount of the spin column.
