PureLink™ HiPure Plasmid Miniprep Kit
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PureLink™ HiPure Plasmid Miniprep Kit
Invitrogen™

PureLink™ HiPure Plasmid Miniprep Kit

The PureLink™ HiPure Plasmid Miniprep Kit allows isolation of high yields of highly pure plasmid DNA from E.coli in lessRead more
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Catalog NumberIncludesNo. of Reactions
K21000225 Preps
K210003100 Preps
Catalog number K210002
Price (USD)
264.00
Each
Add to cart
No. of Reactions:
25 Preps
Price (USD)
264.00
Each
Add to cart

The PureLink™ HiPure Plasmid Miniprep Kit allows isolation of high yields of highly pure plasmid DNA from E.coli in less than 2 hours, without the use of any organic solvents or cesium chloride, and contains low endotoxin levels.
• Fast—Highly-quality purified plasmid DNA in under 2 hours
• Efficient—Increased yields with low endotoxin levels

Plasmid DNA of High Purity
The PureLink™ HiPure Plasmid Miniprep Kit is designed to efficiently isolate plasmid DNA from E.coli in 1.5-2 hours. The kit employs anion-exchange resin composed of small particles with a uniform pore size to purify plasmid DNA to a level equivalent to two passes through CsCl gradients (Figure 1). The need for additional steps to remove contaminants such as phenol, chloroform, and CsCl are eliminated, minimizing exposure to and disposal of hazardous materials.

High Yield and Efficient Endotoxin Removal
PureLink™ HiPure Plasmid Miniprep Kit allows purification of up to 30 μg of high quality plasmid DNA from 1-3 mL overnight E.coli cultures in approximately 1 hour, with an endotoxin level of 0.1-1 EU/ μg, when cloning high copy number plasmids (Table 1). Typically, DNA isolated using the PureLink™ HiPure Plasmid Miniprep Kit has an A260/A280 > 1.80 when samples are diluted in Tris-HCl (pH 7.5) indicating that the DNA is reasonably free of proteins that could interfere with downstream applications.

Isolated DNA can be used for a Number of Downstream Applications:
The purified DNA is ultrapure and suitable for downstream applications, including those requiring the highest purity DNA, such as mammalian transfections (Figure 2), automated and manual DNA sequencing, PCR amplification, in vitro transcription, bacterial cell transformation, cloning, and labeling.

For Research Use Only. Not intended for diagnostic procedures.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
FormatKit
Isolation TechnologyAnion Exchange Resin
Sample TypeBacterial Culture
ScaleMini
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
Prep Scale< 100 μg (Small-Scale) Plasmid DNA
Product LinePureLink™
Product TypePlasmid MiniPrep Kit
Quantity25 Preps
Shipping ConditionRoom Temperature
TargetBAC DNA, Plasmid DNA
Test Time2 hr.
Unit SizeEach
Contents & Storage
• 10 mL Resuspension Buffer (R3)
• 100 μL RNase A
• 10 mL Lysis Buffer (L7)
• 10 mL Precipitation Buffer (N3)
• 50 mL Equilibration Buffer (EQ1)
• 125 mL Wash Buffer (W8)
• 25 mL Elution Buffer (E4)
• 15 mL TE Buffer
• 25 HiPure Columns

Store all components at room temperature.

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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