PureLink™ Pro Quick96 Plasmid Purification Kit
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PureLink&trade; <i>Pro</i> Quick96 Plasmid Purification Kit
Invitrogen™

PureLink™ Pro Quick96 Plasmid Purification Kit

The PureLink™ Pro Quick96 Plasmid Kit combines advanced silica plate extraction chemistry with an optimized 96-well plate design for manualRead more
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Catalog NumberQuantity
K211004A4 x 96 Preps
K211024A
also known as K2110-24A
24 x 96 Preps
Catalog number K211004A
Price (USD)
1,296.00
Each
Add to cart
Quantity:
4 x 96 Preps
Price (USD)
1,296.00
Each
Add to cart

The PureLink™ Pro Quick96 Plasmid Kit combines advanced silica plate extraction chemistry with an optimized 96-well plate design for manual or automated processing of plasmid DNA from E. coli in as little as 45 minutes (Table 1). Using the PureLink™ Pro Quick96 Plasmid Kit, you will:

• Obtain consistently high yields of high-purity plasmid DNA
• Experience ease of use and better performance due to improved plate design and protocol
• Increase processing flexibility with protocols for centrifuge, vacuum manifold, or automated platforms

How it works
The PureLink™ Pro Quick96 Plasmid Kit uses two 96-well plates, a filtration plate (clear O-ring), and a binding plate (red O-ring) to purify plasmid DNA. Bacterial cells are subjected to alkaline lysis and are pelleted by centrifugation. The bacterial lysate is then applied to the Quick96 Filtration Plate for clearing and collection in the Quick96 Binding Plate, where the plasmid DNA binds to the silica membrane. After washing steps to remove contaminants, purified DNA is eluted and ready for use in downstream applications. The entire procedure can be completed in 45 minutes (Figure 1).

Improved plate design
The PureLink™ Pro Quick96 Plasmid Kit provides optimized semiskirted 96-well plates for improved performance and results (Figure 2). The Quick96 Filtration Plate is designed for fast and efficient clearing of the bacterial lysates. The Quick96 Binding Plate offers an increased binding capacity of up to 20 μg plasmid DNA. In addition, the plate nozzles with annular collar prevent cross-contamination of samples and improve drying of the silica membrane to prevent ethanol carryover. In addition, sample processing can be performed manually using a benchtop vacuum manifold, by centrifugation, or with an automated liquid handling platform. The semiskirted plate design is compatible with most vacuum manifolds on robotic workstations.

Higher yields and purity
The PureLink™ Pro Quick96 Plasmid Kit allows rapid and reproducible purification of up to 20 μg of plasmid DNA from E. coli strains grown in up to 5 mL LB, 3 mL 2xYT, or 3 mL TB. In an automated platform using 1.5 mL E. coli culture grown overnight in a square-well block, typical yields are 5-15 μg plasmid DNA, with an average of 9.4 μg (Figure 3). The resulting plasmid DNA is supercoiled with no detectable genomic DNA or RNA (Figure 4). The PureLink™ Pro Quick96 Plasmid Kit provides higher yields of high-quality plasmid DNA for use in common downstream applications such as automated sequencing, PCR, restriction enzyme digestion, and cloning.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Format96-well Plate
Isolation TechnologySilica Plate
Sample TypeBacterial Culture
Elution Volume50 μL (centrifuge), 100 μL (vacuum), 150 µ:L
Final Product TypePlasmid DNA
For Use With (Equipment)Biomek™ FX, Robotic Liquid Handling Systems, Vacuum Manifold, Eppendorf epMotion, Centrifuge, TECAN Freedom Evo, TECAN Genesis
For Use With (Application)Next-Generation Sequencing, Sequencing, PCR, Cloning
High-throughput CompatibilityHigh-throughput Compatible, Not High-throughput Compatible (Manual)
Prep Scale< 100 μg (Small-Scale) Plasmid DNA
Product LinePureLink™
Product TypePlasmid Purification Kit
Quantity4 x 96 Preps
PurityMolecular Biology Grade
Shipping ConditionRoom Temperature
TargetPlasmid DNA
System TypePureLink™
Test Time45 min.
Yield15 μg
Starting Material Amount1.2-1.5 ml lysate⁄well
Unit SizeEach
Contents & Storage
The PureLink™ Pro Quick96 Plasmid Kit includes PureLink™ Pro Quick96 Resuspension Buffer, Lysis Buffer, Neutralization Buffer, RNase A, Wash Buffers, Elution Buffer, square-well blocks, Quick96 Filtration Plates, Quick96 Binding Plates, wash plates, elution plates, and a manual. Store components at room temperature. Store Resuspension Buffer after addition of RNase A at +4°C. Guaranteed stable for one year when properly stored.

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.