*The Agencourt Genfind System formulation has been updated. For more information, please click here.*

The Agencourt Genfind system isolates and purifies genomic DNA (gDNA) from whole blood and serum. Utilizing cutting-edge SPRI® paramagnetic bead-based technology effectively produces a high recovery of DNA for downstream applications such as PCR¹ and genotyping. The process does not require filtration or centrifugation, and supports 96-well plate and tube processing formats.

Application: Genomic DNA isolation from whole blood, plasma, and serum

Downstream Application Techniques: Genotyping applications (SNP, fragment analysis), clinical sequencing, resequencing.

Agencourt Genfind Features Typical DNA recovery of 7 - 12 µg gDNA from 200 µL human blood Processing from 50-400 µL volumes High gDNA recovery Intact, pure gDNA, average size approximately 40 kb Process in 2 mL tubes or 96-well plates No PCR inhibition from common anticoagulants in blood

1. Lyse whole blood or serum in Lysis Buffer and Proteinase K. 2. Bind genomic DNA to paramagnetic beads. 3. Separate beads from contaminants. 4. Wash the magnetic beads with Wash Buffer to remove contaminants. 5. Wash the magnetic beads with 70% Ethanol to remove contaminants. 6. Elute DNA from magnetic particles. 7. Transfer to new plate.

The Agencourt Genfind system efficiently extracts gDNA using flexible starting volumes that can range from 50-200 µL in 96-well format and up to 400 µL in tube format. Figure 1 demonstrates how Agencourt Genfind delivers up to 5 times greater yield in comparison to other technologies.

The Agencourt Genfind process extracts and purifies gDNA of a high molecular weight, producing DNA that is intact and useful for a variety of downstream applications such as cloning or SNP genotyping. Figure 2 represents the quality of the high molecular weight gDNA recovered from human whole blood samples. The average size of the gDNA recovered was approximately 40 kb.

Figure 1: Comparison of gDNA yields from 200 µL of fresh human blood preserved with EDTA using the Agencourt SPRI technology and competitor kits. Results shown are the average and standard deviation from three preparations. All isolations were performed according to manufacturers' protocol. DNA was quantified using the Quant-It2 PicoGreen2 ds DNA Assay kit.

Figure 2: : Pulsed-field gel electrophoresis of gDNA isolated with the Agencourt Genfind purification process. Lane 1: 40 kB T7 DNA size marker. Lanes 2-5: One (1) µg each of four separate Agencourt Genfind preparations of gDNA.

The Agencourt Genfind purification system efficiently removes residual anticoagulants, such as heparin, EDTA, and Citrate. High purity gDNA can be isolated from blood preserved with all of the common anticoagulants with no inhibition of PCR detected (Figure 3).

Figure 3: The Agencourt Genfind kit was used to isolate gDNA from whole horse blood. A) gDNA isolated from 200 µL of horse blood preserved with citrate (lanes 1-3), EDTA (lanes 4-6), or heparin (lanes 7-9), stored at 4°C for less than two weeks. B) PCR of gDNA using transferrin primers. A 1 µL aliquot from each sample shown Figure 3A was used as template in a 10 µL PCR with horse transferrin primers. The expected product size is 389 bp.

The use of the Agencourt SPRI paramagnetic bead-based technology allows Agencourt Genfind to be fully automated while maintaining a high sample quality. Genfind is currently optimized on Beckman Coulter's Biomek® FX, FX^P, NX, and NX^P Laboratory Automation Workstations using either a 96-well multichannel head or a Span-8 head.

To Place an Order or Receive Technical Assistance: In the U.S. and Canada call us at 800-361-7780. Outside the U.S. and Canada call us at 978-867-2600. Email us at webinfo@agencourt.com.

¹ The PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffman-La Roche, Ltd.
² All trademarks are property of their respective owners.