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NucleoFast 96 PCR

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Cost-efficient 96-well ultrafiltration kit for PCR clean-up

For Purification of PCR products > 150 bp


Ready-to-use DNA for sequencing and microarray spotting
• Sturdy membrane allows easy recovery of DNA
• No well-to-well cross-contamination
• Fast procedure
• Available as complete kits and as plates only
• Detergent-free membrane
• Manual or automated use

The NucleoFast® technology is based on an effective removal of contaminants by means of ultrafiltration, thus eliminating the need for chaotropic salt for binding and ethanolic wash buffers.
During the NucleoFast® 96 PCR procedure the PCR sample (up to 300 μL) is applied to the ultrafiltration membrane. Under vacuum or centrifugation contaminants (primers, dNTPs, salts) are filtered to waste. The PCR products can be recovered from the membrane after the addition of water or buffer and a short incubation (1–3 minutes) and mixing step. PCR products are ready-to-use for downstream applications (e.g., sequencing and microarray analysis).


NucleoFast® is available in the convenient kit format, including the NucleoFast® 96 PCR plates, necessary buffers, and consumables. The NucleoFast® 96 PCR plates are also available separately to offer a greater flexibility.

Manual and automated high throughput (HTP) kits

  • NucleoFast® 96 PCR Clean-up Kit
  • NucleoFast® 96 PCR Plates
Characterization:

 

The NucleoFast® technology is based on an effective removal of contaminants by means of ultra filtration, thus eliminating the need for chaotropic salt for binding and ethanolic wash buffers.

 

NF-96-PCR

During the NucleoFast® 96 PCR procedure the PCR sample (up to 300 μL) is applied to the ultrafiltration membrane. Under vacuum or centrifugation contaminants (primers, dNTPs, salts) are filtered to waste.The PCR products can be recovered from the membrane after the addition of water or buffer and a short incubation (1–3 minutes) and mixing step. PCR products are ready-to-use for downstream applications (e.g., sequencing and microarray analysis).
Technology Ultrafiltration technology
Format 96-well plates
Processing Manual or automated, vacuum or centrifugation
Sample material 20–300 µL
Fragment size > 150 bp
Typical recovery 40–95%
A260/A280 1.70–1.80
Recovery volume 25–100 µL
Preparation time 20 min/plate (for typical PCR reactions of 25 µL)


Technical Data:

Appplication data
Recovery rate depends on length of PCR product
NucleoFast® 96 PCR shows a high recovery even for small fragments
 

 

 

 

Complete removal of primers
PCR products (164 bp, 252 bp) were amplified with an excess of primers. PCR products (20 μL) were then purified manually using NucleoFast® 96 PCR (recovery volume: 100 μL), dried, resuspended in 15 μL loading buffer, and analyzed on a 1% TAE agarose gel. Small fragments are recovered effectively whereas primers are completely removed.
C: Unpurified control
M: Marker
Reliable recovery of PCR products
PCR products (300–350 bp) were amplified in a buffer containing either 100 mM betaine or 20% Q-Solution (Qiagen) and have been purified manually using NucleoFast® 96 PCR under vacuum according to the standard protocol including the optional washing step with water. Recovery has been compared to NucleoSpin®Multi-96 Extract. M: marker.
Data kindly provided by Dr. Silvia Rüberg and Eva Schulte-Berndt, Center for Genome Research, Transcriptomics, University of Bielefeld, Germany
PCR products purified with NucleoFast® 96 PCR are suitable for direct microarray spotting
96 PCR products (PCR on bacterial cultures) have been purified manually using NucleoFast® 96 PCR with an additional washing step with water before recovery of DNA. PCR products were not normalized for concentration and were spotted with Telechem Splitpin, 2xSSC. PCR products were then hybridized with labeled cDNA (Cy5 – left panels, Cy3 – right panels).
Data kindly provided by Dr. Jürgen Zimmermann,
Genome Core Facility, EMBL, Germany

High recoveries and consistencies
PCR products with different lengths (164 to 1484 bp) have been purified with either NucleoFast® 96 PCR or product M, based on ultrafiltration technology (12 replicates per purification kit, starting from one PCR pool per PCR product). NucleoFast® 96 PCR shows a higher amount of DNA after the purification, thus higher recovery, and a higher consistency.
Reliable fully-automated rapid purification of PCR products
96 PCR products of different lengths (164, 252, 359, 459, 645, 787, 982, and 1484 bp, 25 μL each) have been purified fully automated using NucleoFast® 96 PCR on a Biomek® FX (Beckman-Coulter) according to the standard protocol and were analyzed on a 1% TAE agarose gel (M: marker). Recovery of fragments from the membrane was facilitated by 5 mixing cycles and a subsequent 1 minute incubation after addition of recovery buffer. Total preparation time for 96 samples is about 15 minutes.
Reliable fully-automated clean-up of PCR products
PCR fragments of different lengths (25 μL each) have been purified fully automated on a Tecan Genesis™ 150 using NucleoFast® 96 PCR according to the standard protocol and were analyzed on a 1% TAE agarose gel (control: unpurified samples, M: marker). Recovery of fragments from the membrane was facilitated by 5 mixing cycles and a subsequent 2 minutes incubation after addition of buffer
(5 mM Tris/HCl pH 8.5).


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