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Table 4 Incubation at 65 °C for 1 h is superior to overnight incubation for recovery of parasite DNA from dried blood spots

From: An improved nucleic acid extraction method from dried blood spots for amplification of Plasmodium falciparum kelch13 for detection of artemisinin resistance

Incubation time at 65 °C 1 h Overnight
Lysis buffer 3 M GuSCN 3 M GuSCN + 16.7% ISOH 3 M GuSCN 3 M GuSCN + 16.7% ISOH
Average Ct for Pf 18S rDNA (± SD) Undetectable 30.0 (± .3) Undetectable Undetectable
Average Ct for human actin (± SD) 29.8 (± .4) 28.3 (± .3) 28.4 (± .3) 26.5 (± .9)
  1. Experiments done with samples at parasite density of 2000 parasites/mL
  2. Pf, Plasmodium falciparum; GuSCN, guanidine thiocyanate; ISOH, isopropanol; SD, standard deviation; Ct, cycle threshold