High-speed shaking method for DNA extraction from frozen blood clots
Author Information
Author(s): Klara Lundblom, Alex Macharia, Marianne Lebbad, Adan Mohammed, Anna Färnert
Primary Institution: Karolinska Institutet
Hypothesis
Can high-speed shaking improve DNA extraction from frozen blood clots compared to traditional methods?
Conclusion
High-speed shaking is an effective method for homogenizing frozen blood clots, resulting in high-quality DNA suitable for PCR analysis.
Supporting Evidence
- High-speed shaking achieved the highest sensitivity for PCR detection of malaria parasites.
- The method generated the highest DNA yield compared to commercial protocols.
- PCR detection levels were similar to those obtained from blood collected with anticoagulants.
- High-speed shaking reduced the risk of cross-contamination during sample processing.
- 92% success rate in genotyping the human thalassaemia gene using the new method.
Takeaway
Shaking frozen blood clots really fast helps get better DNA for testing, which is important for finding diseases like malaria.
Methodology
The study evaluated high-speed shaking of blood clots for DNA extraction and compared it with two commercial methods.
Potential Biases
Manual handling in traditional methods poses a risk of cross-contamination.
Limitations
A systematic assessment of different clot protocols in field samples could not be performed due to lack of additional clots from the same individuals.
Participant Demographics
Participants included asymptomatic children aged 0-2 years in Kenya.
Digital Object Identifier (DOI)
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