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

PureLink™ HiPure Plasmid Filter Midiprep Kit

The PureLink™ HiPure Plasmid Filter Midiprep Kit is designed to isolate plasmid DNA of the highest purity from 25 toRead more
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Catalog NumberQuantity
K21001425 Preps
K21001550 Preps
Catalog number K210014
Price (USD)
388.00
Each
Add to cart
Quantity:
25 Preps
Price (USD)
388.00
Each
Add to cart

The PureLink™ HiPure Plasmid Filter Midiprep Kit is designed to isolate plasmid DNA of the highest purity from 25 to 100 mL of an overnight bacterial culture. The PureLink™ HiPure Plasmid Filter Kits include HiPure Filter Columns, which combine filtration for clarification of the bacterial lysate with anion-exchange resin purification of plasmid DNA, in a single unit. The resulting plasmid DNA is of a quality equivalent to that obtained by purification with two passes through cesium chloride gradients—the most rigorous method for DNA purification. Purify high-quality DNA in less than two hours for transfections and other applications with no need for additional steps to remove contaminants like RNA, proteins, or endotoxins. Additionally, the elimination of phenol, chloroform, ethidium bromide, and cesium chloride from the protocol minimizes exposure to, and disposal of hazardous materials. The PureLink™ HiPure Plasmid Filter Kits provide:

• Simple protocols—bacterial lysate clarification without centrifugation
• Fast results—save time by purifying plasmid DNA in less than two hours
• High yields—up to 200 μg for midipreps and up to 850 μg for maxipreps (see table)
• Reliable performance—the high-quality plasmid DNA purified with these kits is suitable for a wide variety of applications

How it works
The addition of a HiPure Filter Column to the PureLink™ HiPure Plasmid Kits provides rapid clearing of the bacterial lysate without the need for centrifugation (see figure). The lysate filtration cartridge is integrated into the DNA-binding column for a combined step of lysate clarification and plasmid DNA binding directly to the anion-exchange resin. A single column wash removes impurities such as RNA, proteins, metabolites, and other low molecular weight molecules; the ultrapure plasmid DNA is then eluted with a high-salt buffer. The DNA is then desalted and concentrated with an isopropanol precipitation step, then collected by centrifugation. The entire protocol can be completed in about two hours.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Column TypeFilter Column
FormatColumn
Isolation TechnologyAnion Exchange Resin
Sample TypeBacterial Culture
Elution Volume5 mL
Final Product TypePlasmid 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 Filter MidiPrep Kit
Quantity25 Preps
PurityTransfection Grade
Shipping ConditionRoom Temperature
TargetPlasmid DNA
System TypePureLink™
Test Time90 min.
Yield350 μg
Unit SizeEach
Contents & Storage
The PureLink™ HiPure Plasmid Filter Kits include Equilibration Buffer (EQ1), Resuspension Buffer (R3), Lysis Buffer (L7), Precipitation Buffer (N3), RNase A, Wash Buffer (W8), Elution Buffer (E4), TE Buffer (TE), and HiPure Filter Columns. Store all components at room temperature upon receipt.

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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