GenFind V3 Performance and Data
Genomic DNA Extraction from Blood and Serum
The GenFind V3 kit is a genomic DNA (gDNA) isolation reagent kit built on SPRI paramagnetic bead-based technology. It enables purification of PCR compatible gDNA from fresh and frozen blood collected in tubes containing anticoagulants such as EDTA, citrate or heparin. The method can be run manually in a 2 mL tube format or 96-well format, or automated in 96-well format on variety of Beckman Coulter Biomek liquid handling workstations. Consistent recovery of gDNA from various types of fresh and frozen blood samples compared to other suppliers.
- Optimal and consistent purity and quality of gDNA isolates
- No carryover of inhibitors from various anticoagulants for exceptional PCR and other down stream assays
- Process up to 400 µL of fresh or frozen blood
Competitive Recovery and Purity of gDNA
Genomic DNA was extracted from 200 µL of fresh or frozen blood collected in tubes containing heparin, EDTA or Citrate with GenFind V3 and other suppliers’ kits. In Figure 1 samples were quantitated with the Quant-iT DNA assay kit, Picogreen, and NanoDrop (Thermo Fisher Scientific) for DNA yield. Higher amounts of gDNA were recovered using the GenFind V3 kit over other suppliers’ kits in one type of blood collection tube and competitive yields were recovered from the other two tube types. In Figure 2 samples were accessed for purity using the NanoDrop (Thermo Fisher Scientific). For all types of fresh and frozen blood samples, GenFind V3 purified gDNA with satisfactory A260/280 ratios.
Enhanced Purity of gDNA with GenFind V3Genomic DNA was extracted from 200 µL of fresh or frozen blood collected in tubes containing heparin, EDTA and citrate with GenFind V3, GenFind V2 and three other suppliers’ kits. Samples were accessed for purity using the NanoDrop (Thermo Fisher Scientific). For every tube type, GenFind V3 extracted gDNA with a greater purity than GenFind V2 and the other supplier’s kits.
Increase Yield with Increase of Input Volume of Blood
Increasing the volume of the input can increase total yields without affecting purity. Genomic DNA was extracted from 400 µL of fresh blood collected in tubes containing EDTA with GenFind V3. Samples were accessed for purity and yield using the NanoDrop (Thermo Fisher Scientific).
No Carryover of Inhibitors for Exceptional PCR and Downstream Applications
The ability to PCR was assessed via qPCR using a primer set (forward primer 5´-ggacttcgagcaagagatgg-3´ and reverse primer 5´-agcactgtgttggcgtacag-3´) designed to span Exon 4 and 5 of the beta (β)-actin gene (ActB) to produce 327 base pair amplicons. In Figure 6 the amplification plot of 10ng of DNA extracted from tubes containing citrate, EDTA or heparin. For all tube types we get amplification of DNA indicating that there are no carryovers of PCR inhibitors for any of the anticoagulants. In Figure 7 the amplification plot of 10ng of DNA extracted from frozen tubes containing citrate using GenFind V3 and GenFind V2.
High Recovery of High Molecular Weight DNAGenFind V3 isolates high quality gDNA. Genomic DNA isolated from various blood collected in heparin, EDTA and citrate tubes was run on the Agilent Genomic DNA ScreenTape to assess quality. In the image to the right DIN values from all samples isolated with GenFind V3 were all ≥ 9.0, indicating that high quality and intact gDNAs were recovered. DNA isolated using Supplier A’s kit were of lower quality indicated with a DIN score of 7.8. In both Figure 9 and Figure 10 sample traces of the Genomic DNA isolated from tubes containing heparin and citrate.
1. Lyse whole blood or serum in Lysis Buffer and Proteinase K
2. Bind DNA to magnetic beads
3. Separate beads from contaminants
4. Wash the magnetic beads with Wash Buffer 1 to remove contaminants
5. Wash the magnetic beads with Wash Buffer 2 to remove contaminants
6. Elute DNA from magnetic beads
7. Transfer to new plate for storage
Less Hands-on-Time with Less Pipette Actions
Figure 11 represents hands on time to extract gDNA for 1 to 96 samples using GenFind V3 or a column based supplier. Even at 10 samples hands on time to extract DNA from blood is faster using GenFind V3. In Figure 12 the total number of pipette actions required for 1, 8, 24, 48, and 96 samples. Pipette actions include dispensing the sample, mixing the sample and discarding the supernatant. With the ability to use a multichannel pipette there is less pipette actions that need to take place at 24 or more samples than with column based suppliers.
Products for DNA Isolation from Blood
GenFind V3 Reagent Kit is available in 2 kit sizes based on your throughput needs. Contact your local sales representative or visit our product page to request a quote.