PureLink™ HiPure Plasmid Midiprep Kit
PureLink™ HiPure Plasmid Midiprep Kit
Invitrogen™

PureLink™ HiPure Plasmid Midiprep Kit

The PureLink HiPure Plasmid Midiprep Kit is designed to isolate transfection-grade plasmid DNA from E. coli.
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Catalog NumberQuantityIncludes
K21000425 Preps
K21000550 Preps
Catalog number K210004
Price (EUR)
256,00
Each
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Quantity:
25 Preps
Price (EUR)
256,00
Each
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The PureLink™ HiPure Plasmid Midiprep Kit is designed to isolate transfection-grade plasmid DNA from E. coli. The Midiprep Kit protocol will typically yield 100–350 μg plasmid DNA from 15–25 mL bacterial culture, at a purity that is comparable to that achieved by two passes through cesium chloride gradients.

Advantages of the PureLink™ HiPure Plasmid Midiprep Kit include:

  • Labor and time saving—no need for extra steps to remove endotoxin and other contaminants
  • High-purity, low-endotoxin product—pure enough for mammalian cell transfection
  • Versatility—purify any type and size of plasmid DNA, including BAC, bacmids, and ssM13 DNAs

Anion-exchange chromatography for plasmid purification

The PureLink™ HiPure Plasmid DNA Purification Kits use a patented anion-exchange resin to purify plasmid DNA to a level equivalent to two passes through CsCl gradients. The resin combines excellent capacity with a fast flow rate, high resolution, high yield, and efficient endotoxin removal. Plasmid preparations can typically be completed in less than 2 hours.

Contaminant-free DNA

Unlike cesium chloride gradient protocols, PureLink™ HiPure Plasmid Purification Systems do not use organic solvents, ethidium bromide, or cesium chloride, all of which are troublesome to work with and to dispose of. Typically, plasmid DNA prepared using the PureLink™ HiPure Plasmid Midiprep Kit will have an A260/A280 ratio of greater than 1.80, indicating that the DNA is reasonably free of proteins that might interfere with downstream applications. Endotoxin levels are typically within 0.1–1 EU/μg, making this plasmid DNA ideal for mammalian cell transfection.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Elution Volume5 mL
Final Product TypeBAC DNA, Plasmid DNA
For Use With (Application)Next-Generation Sequencing, Transfection, Cloning, Sequencing, Transformation, Nucleic Acid Labeling, PCR, In Vitro Transcription
High-throughput CompatibilityNot High-throughput Compatible (Manual)
No. of Reactions25 Preps
Plasmid<40kb, Low Copy Plasmid, High Copy Plasmid, BAC
Prep Scale100-200 μg (Medium-Scale) Plasmid DNA
Product LinePureLink™
Product TypePlasmid MidiPrep Kit
PurityTransfection Grade
Quantity25 Preps
Sample TypeBacterial Culture
Shipping ConditionRoom Temperature
System TypePureLink™
TargetBAC DNA, Plasmid DNA
Test Time2 hr.
Yield100 to 350 μg
FormatColumn
Isolation TechnologyAnion Exchange Resin
Unit SizeEach
Contents & Storage
• 100 mL Resuspension Buffer (R3)
• 550 μL RNase A
• 100 mL Lysis Buffer (L7)
• 100 mL Precipitation Buffer (N3)
• 250 mL Equilibration Buffer (EQ1)
• 500 mL Wash Buffer (W8)
• 125 mL Elution Buffer (E4)
• 15 mL TE Buffer
• 25 HiPure Columns
• 5 Column Holders

Store all components at room temperature.
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Frequently asked questions (FAQs)

I'm getting low/no plasmid DNA after purification using a PureLink HiPure kit, even though there was measurable absorbance. Do you have any suggestions for what I can do?

A common problem encountered with absorbance measurements is turbidity of samples. (This could be caused by residual resin from the column.) If there is insoluble material in the cuvette (not often detected by the naked eye), much of the UV light is not absorbed but scattered, leading to an artificially high UV absorbance reading (at 260 or 280 nm, for example.) If your A260 is high, we recommend that you check the A320 to determine if there is resin in the sample. You can also try to centrifuge or filter (0.2 µm filter) your sample to remove any resin and then recheck the concentration.

I've run out of buffer when using the PureLink HiPure Plasmid Purification Kit (Cat. No. K210018). Can I purchase the buffers separately?

Yes, we would recommend purchasing the PureLink HiPure BAC Buffer Kit (Cat. No. K210018). This kit includes Resuspension Buffer (R3) (250 ml), Lysis Buffer (L7) (250 ml), Precipitation Buffer (N3) (250 ml), and RNase A (20 µg/ml) (5 ml).
You will need to add less RNase A than stated on the bottle label of the R3 buffer in this kit. It says to add 5.6 mL of RNase A. This is the correct amount for the BAC protocol; however, if you are performing standard plasmid isolation, 1.4 mL RNase A should be added.

Plasmid DNA isolated using a PureLink column-based purification kit from an endA+ strain is degraded after a restriction digest. Do you have a suggestion for this?

The HiPure kits should remove all protein from the DNA including endonucleases. For the silica-based PureLink Quick Plasmid Miniprep Kit, we recommend an extra wash with the optional Wash Buffer W10 to remove endonucleases. This solution is not compatible with the HiPure system and should not be used with those kits. Alternatively, heat the eluted DNA in TE for 10 min at 70 degrees C. This should heat-inactivate any contaminating nucleases.

I'm seeing extra bands present after plasmid purification using your PureLink column-based system. What could cause this to happen?

Extra bands can occur when plasmid DNA is nicked and/or permanently denatured. Plasmid DNA that has been nicked (covalently opened) will run slower than supercoiled DNA during electrophoresis. A small amount of this species of DNA is common and is suitable for downstream applications. Permanently denatured DNA will migrate ahead of the supercoiled DNA and may not be suitable for downstream applications. Do not allow the lysis reaction to proceed longer than 5 minutes.

My purified DNA has particles in it after column-based plasmid purification. Any suggestions?

We have seen this on occasion. The particles do not affect quality of the DNA. Remove the particles by performimg a 1 minute centrifugation at 12,000 x g.

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3270056Certificate of AnalysisJun 27, 2025K210005
3269502Certificate of AnalysisJun 26, 2025K210004
3248239Certificate of AnalysisMay 28, 2025K210005
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