Extracting pure DNA from whole blood is challenging. Mammalian blood has few DNA-containing cells and the large amounts of haemoglobin and plasma proteins can clog up extraction matrices and inhibit downstream applications. There is always a risk that blood samples contain harmful infectious agents. GeneMole® is a new bench-top automated system for extraction of high quality nucleic acids from many different sources, including blood. The system processes up to 16 samples/run; 8 samples in 28 min. GeneMole® handles sample volumes ≤200 µl. Reagents are pre-filled in individually sealed strips (MoleStrips™) for maximum convenience and safety.
Materials
GeneMole® Instrument
(Mole Genetics AS, product no. MG10-000)
MoleStrips™ DNA Blood
(Mole Genetics AS, product no. MG10-101)
Barrier Tips 1250 µl
(VWR, product no. VWR 732-0908)
Tube MTS 8-strip 1.1 ml Rack Sterile
(Axygen, product no. MTS-11-8-C-R-S)
Tube 8-strip CAP MTS Sterile
(Axygen, product no. MTS-8CP-C-S) Methods Extraction Set-up (3 min)
Load MoleStrips™ DNA Blood, pipette tips, and elution tubes onto the freestanding work tray according to the chosen number of samples (1–16).
Add 100 or 200 µl whole blood to each of the sample tubes. Blood from birds or samples with very high white cell counts should be diluted in PBS.
Place the sample tubes on the work tray, and then the work tray into GeneMole®.
Choosing Program (1 min)
Select the program “DNA from blood” from the touch screen.
Choose input and elution volume from the drop-down menu.
Start the instrument.
DNA Extraction (1-8 samples, 28 min; 9-16 samples, 60 min)
Extraction of DNA from blood on GeneMole® is a completely automated process.
Lysis buffer is added to the blood.
DNA is bound to magnetic beads. An external magnet is used to separate the DNA-bead complex from the remaining sample.
Two gentle, yet efficient washing steps, secure removal of haemoglobin and other inhibitors.
Purified DNA is finally eluted in Tris-HCl buffer.
Results
The yield and quality of the extracted DNA (measured by NanoDrop® or standard spectrophotometer) are listed in the table. High quality DNA was obtained from all the tested samples.
Automated DNA extraction gave highly reproducible results without any visible cross-contamination (Figure 1). The DNA was free from inhibitors and compatible with a range of downstream analysis methods.
Figure 1. DNA extraction with GeneMole® gives highly reproducible results (CV <5%) (Click to enlarge)
Table 1. Yield and Quality of Genomic DNA Extracted from Blood from Different Species by GeneMole® (Click to enlarge)
Correspondence
Address correspondence to Mole Genetics AS, Volls vn. 13D, 1366 Lysaker, Norway; Tel.: +47 67 10 18 20; [email protected], www.molegenetics.com.
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