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A Practical and Novel Method to Extract Genomic DNA from Blood Collection Kits for Plasma Protein Preservation
Published on: May 18, 2013
Plasma preparation from whole blood using ultrasound
C M Cousins1, P Holownia, J J Hawkes
1School of Biosciences, Cardiff University, Main Building, Park Place, Wales CF1 3TL., Cardiff, UK.
Abstract:
A technique to efficiently separate plasma from human whole blood is described. Essentially, 3-mL samples are held on the axis of a tubular transducer and exposed for 5.7 min to an ultrasonic standing wave. The cells concentrate into clumps at radial separations of half wavelength. The clumps grow in size and sediment under gravity. A distinct plasma/cell interface forms as the cells sediment. The volume of clarified plasma increases with time. The separation efficiencies of transducers of 29-mm and 23-mm internal diameters driven, by test equipment, at radial resonances of 3.4 and 1.5 MHz, respectively, were compared. The average efficiency of separation was 99.6% at 1.5 MHz and 99.4% with the 3.4-MHz system. The cleared plasma constituted 30% of the sample volume at 1.5 MHz and 25% at 3. 4 MHz. There was no measurable release of haemoglobin or potassium into the suspending phase, indicating that there was no mechanical damage to cells at either frequency. A total of 114 samples from volunteers and patients were subsequently clarified in a 1.5-MHz system driven by an integrated generator. The average efficiency of clarification of blood was 99.76% for the latter samples. The clarification achieved is a significant improvement on that previously reported (98.5%) for whole blood exposed to a planar ultrasonic standing wave field (Peterson et al. 1986). We have, therefore, now achieved a six-fold reduction of cells in plasma compared to previous results.
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