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The PureLink® Pro 96 Viral RNA⁄DNA Purification Kit is the only 96-well silica plate system that can extract both viral RNA and DNA from 200-µl cell-free fluids which uses a low-depth 96-well centrifuge bucket for ease-of- use, all without any loss in sensitivity. These kits provide: • Versatility-one kit for all your samples, the protocol is optimized for both viral RNA and DNA extraction • Flexibility in centrifuge choice-minimum speeds for 96 well processing of 2250 x g and a rotor bucket depth of 5 cm make this kit one of the most adaptable systems for existing laboratory centrifuges (Figure 1) • Sensitivity and reproducibility as good as a spin column-moving to high throughput is reliable and easy with no loss in accuracy of quantitation
The PureLink® Pro 96 Viral RNA⁄DNA Purification Kit is based on conventional silica plate extraction chemistry. Cell-free samples are lysed in guanidine containing buffers that have been reformulated to increase the recovery and sensitivity of purified nucleic acids, while carrier RNA is used to protect samples from RNase degradation. Binding of lysates to the Viral Filter Plate can be performed with either a vacuum manifold or centrifugation using a 96-well rotor with buckets having a depth of only 5 cm. After washes with ethanol-based buffers, the viral RNA or DNA is eluted in 100 µl of RNase-free water. The RNA or DNA is ready to use in one-step or two-step RT-PCR and qRT-PCR, qPCR, or other enzymatic amplification procedures.
The PureLink® Pro 96 Viral RNA⁄DNA Purification Kit shows excellent well-to-well consistency A critical requirement for any 96-well purification system is well-to-well consistency. When using a technique as sensitive as qRT-PCR for viral nucleic acid detection, which may exist as low viral titers from patient samples, consistency between samples becomes even more important. When well-to-well recovery of lentiviral RNA was analyzed using the PureLink® Pro 96 Viral RNA⁄DNA Purification Kit, consistent Ct values were obtained in qRT-PCR analyses while plasma controls run on the same plate were negative, showing no cross contamination (Figure 2).
Get the same results in a 96-plate extraction as you would with a spin column To prove consistency of our novel purification chemistry in the both high- and low-throughout formats, Armored RNA® HCV virus was spiked in human plasma and 200 µl were used for viral nucleic acid purification with either the PureLink® Viral RNA⁄DNA Mini Kit (spin-column based purification) or with the PureLink® Pro 96 Viral RNA⁄DNA Purification Kit. The purified samples were analyzed for performance comparison using qRT-PCR. Both purification methods successfully demonstrated viral purification and detection as both purification profiles exhibited scalable qRT-PCR detection analysis profiles (Figure 3). The PureLink® Pro 96 Viral RNA⁄DNA Purification Kit provides sensitive linear scalable viral RNA⁄DNA purification with a sensitivity and efficiency equal to that of the spin column purification format.
Improved plate design The PureLink® Pro Viral RNA⁄DNA purification kit provides optimized semi-skirted 96-well plates (Figure 4) for improved performance and results. 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 (at 2250 x g), or with an automated liquid handling platform. The semi-skirted plate design is compatible with most vacuum manifolds on robotic workstations.
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