PureLink™ 96 HQ Plasmid DNA Purification Kit
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Invitrogen™

PureLink™ 96 HQ Plasmid DNA Purification Kit

The PureLink HQ 96 Plasmid DNA Purification Kit isdesigned for high-throughput isolation of high-qualityplasmid DNA from bacterial cells. It canRead more
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Catalog number K210096
Price (USD)
1,980.00
Each
Add to cart
Price (USD)
1,980.00
Each
Add to cart
The PureLink HQ 96 Plasmid DNA Purification Kit isdesigned for high-throughput isolation of high-qualityplasmid DNA from bacterial cells. It can beused with a vacuum manifold or a centrifuge and iscompatible with automated liquid handling workstations.

The PureLink HQ 96 Plasmid DNA Purification Kit offersthe following advantages:
• Ability to isolate up to 10 μg plasmid DNA per well
• Designed to isolate high-quality plasmid DNA in30-45 minutes
• Minimal genomic DNA contamination in the purifiedplasmid DNA
• Compatible with automated liquid handlingworkstations
• Reliable performance of the purified plasmid DNA indownstream applications such as mammaliantransfection

Process overview
Bacterial cells are harvested and resuspended in ResuspensionBuffer with RNase. The cells are lysed using an alkaline/SDScell lysis procedure. The lysate is then neutralized andconditions are adjusted for subsequent binding. After lysateclarification using the PureLink HQ 96 Clarification Plate, thelysate is processed through the PureLink HQ 96 BindingPlate. The DNA binds to the silica-based membrane in theplate and impurities are removed by washing with WashBuffer. The DNA is then eluted in low salt Elution Buffer orwater.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Format96-well Plate
Isolation TechnologySilica Plate
Sample TypeBacterial Culture
Final Product TypePlasmid DNA
For Use With (Equipment)Automated Liquid Handling Workstation
For Use With (Application)PCR, Transfection, Sequencing, Transformation, Cloning
High-throughput CompatibilityHigh-throughput Compatible
No. of Reactions4 x 96 Preps
Prep Scale< 100 μg (Small-Scale) Plasmid DNA
Product LinePureLink™
Product TypePlasmid DNA Purification Kit
Quantity4 x 96 preps
Shipping ConditionRoom Temperature
TargetPlasmid DNA
Test Time30 to 40 min.
Unit SizeEach
Contents & Storage
Sufficient reagents are provided in the kit to perform 384 (4 x 96) isolations.

Reagents include:
• Resuspension Buffer (R1)
• Lysis Buffer (L1)
• Neutralization/Binding Buffer (B1)
• Wash Buffer (W1)
• Elution Buffer; 10 mM Tris-HCl, pH 8.5 (E1)
• RNase A
• PureLink HQ 96 Clarification Plate
• PureLink HQ 96 Binding Plate
• Receiver Plate
• Adhesive Foil for plates

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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