DNA Extraction Protocol from Dried Blood, Body Fluids and Sperm Spots

Dried blood, body fluids, and sperm samples on filter paper can be processed using the following method. We recommend using specimen paper for spotting blood, as this unique filter paper disintegrates when incubated in aqueous collected using other specimen collection papers.

• Cut or punch out the blood (or other sample) spot from the filter paper. Tear or cut the filter into small pieces and place it into a 1.5- or 2.0-mL centrifuge tube.
• Add 200 μl Buffer MTL to 1-4 punched filter paper circles (3 mm). Followed by the addition of 10 μl Proteinase K solution. Incubate mixture at 55°C for 45-60 minutes. Mix the samples several times during incubation by vertexing.
• Add 220 μl Buffer MSL, close the lid, and mix thoroughly by vertexing for 20 sec at maximum speed.
• Briefly centrifuge the centrifuge tube to remove any liquid droplets from inside the lid.
• Transfer 400 μl supernatant into a new 1.5 mL tube or 96 2-ml deep well plate.
• Add 300 μl absolute ethanol and 10 μl Mag-Bind Particles Solution C to the lysate. Mix thoroughly by shaking at maxi speed for 2 min or pipetting up and down 50 times. Absolute ethanol and Mag-Bind Particles Solution C can be premixed.
• Place the tube/plate on a magnetic separation device to magnetize the magnetic particles. Incubate at room temperature for 10 minutes. Completely remove and discard the cleared supernatant.
• Remove the tube/plate containing the Mag-Bind particles from the magnetic separation device. Add 600 μl Buffer MP/ethanol mixture to each sample. Resuspend the Mag-Bind particles pellet by shaking at max speed for 2 min or pipetting up and down 30 times.
• Place the tube or plate onto the magnetic separation device to magnetize the Mag-Bind particles. Completely remove and discard the cleared supernatant.
• Remove the tube containing the Mag-Bind particles from the magnetic separation device. Add 60 μl SPM Buffer to each sample. Resuspend the Mag-Bind particles pellet by shaking at max speed for 1 min or pipetting up and down 30 times.
• Place the tube onto the magnetic separation device to magnetize the magnetic particles. Completely remove and discard the cleared supernatant.
• Leave the tube to air dry on the magnetic separation device for 10 minutes. Remove any residue liquid with a pipette.
• Remove the tube from the magnetic separation device. Add 100-200 μl Elution Buffer to each sample. Resuspend the Mag-Bind particles pellet by shaking at max speed for 2 min or pipetting up and down 30 times.
• Incubate 10 minutes at 65°C.
• Resuspend the Mag-Bind particles by shaking at max speed for 1 min or pipetting up and down 30 times.
• Place the tube onto a magnetic separation device to magnetize the Mag-Bind particles.
• Transfer the cleared supernatant containing purified DNA to a new plate.

Creative Bioarray Relevant Recommendations

• The biobank of Creative Bioarray is an advocacy-owned repository for biological samples and clinical data. It contains tissue banks, blood samples, RNA samples, DNA samples, and body fluids.
• We provide a series of nucleic acid extraction kits for a variety of samples to help our customers accelerate their research. Beyond that, we also offer extraction services of nucleic acid from a wide range of starting materials.

• Use 1-4 punched circles (3 mm diameter) for each DNA isolation.
• Before starting, bring frozen samples and protease solution to room temperature, and preheat an aliquot of elution buffer (approximately 0.5 mL per sample) at 65°C.

Blood

Biofluids

Nucleic Acid Extraction