Malaria parasite deoxyribonucleic acid (DNA) extraction from dried blood spots (DBS) using QIAGEN QIAMP DNA extraction mini-kit

Two pieces of 3 mm disk from the Whatman filter paper dry blood spots were punched out using a sterile hole punch and placed into appropriately labeled 1.5 micro-centrifuge tube, to this, 180 µl of animal tissue lysis buffer was added to ensure the pieces of filter paper were soaked before incubation at 85 °C for 10 min followed by addition of 20 µl of proteinase K stock solution. The mixture was vortexed and incubated at 56 °C for 1 h after which lysis buffer (buffer AL) was added to the sample, thoroughly mixed by vortexing and incubated at 70 °C for 10 min. Absolute ethanol (200 μl) was added and thoroughly mixed. The mixture was then added to a amp Mini kit spin column placed in a 2 ml collection tube and centrifuged at 8000 rpm for 1 min. The spin column was removed and placed in a clean 2 ml well labeled collection tube while the filtrate was discarded with the tube, 500 μL of Wash buffer (Buffer AW1) was then added and the mixture centrifuged at 8000 rpm for 1 min. The collection tube containing the filtrate was discarded. Buffer AW2 (500 μl) was added to the spin column and then centrifuged at full speed (20,000 × g or 14,000 rpm) for 3 min. Again, the filtrate was discarded and the spin column was placed in a 1.5 mL microcentrifuge tube. DNA of the malaria parasite was eluted with 150 μl elution buffer AE, incubated at room temperature (28 ± 2 °C) for 1 min and centrifuged at 6000 × g (8000 rpm) for 1 min. The extracted DNA was stored in the refrigerator at − 20 °C until it was needed for subsequent molecular studies (Bioscience 2015). The quality and quantity of extracted DNA yield was 1.87 and 160 ng/μL respectively.