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Schizonts and DNA in buffer solution were transferred to an electroporation cuvette and schizonts were transfected using the Amaxa gene pulser and program U33 (Amaxa Biosystems).
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Then, the buffer solution was transferred into the anaerobic chamber.
Each serum sample was initially diluted 1 50 in buffered saline and 50 uL of this solution were transferred into vinyl microplates containing 96 wells (Corning Incorporated, Corning, NY, USA).
To elute bound phage, our standard protocol is to use a citrate buffer (pH 2) for 5 minutes after which the solution is transferred to an Eppendorf tube containing Tris-buffer to ensure an immediate neutralisation of the low pH.
Then, 2.5 μg/ml calcein AM was added to 20 μl DMSO, and 10 μl of this solution was transferred to 12 ml Hanks Buffered Saline Dispense.
A 10-μl aliquot of this solution was transferred to an Eppendorf tube and mixed with a corresponding buffer solution (115 μl).
It was dissolved in lysate buffer and proteinase K, and after another precipitation with ethanol, the solutions were transferred to spin columns (NucleoSpin Tissue).
It was dissolved in lysate buffer and proteinase K, and after a second precipitation with ethanol, the solutions were transferred to spin columns.
The extracted solutions were transferred to PFA centrifuge tubes and 25 mL ammonium-acetate buffer (0.5 mol L−1, pH 5) and 0.2 mL 5% NaBEt4 solution were added.
After being dispersed in buffer solution, sample was transferred into the prefabricated solid-phase extraction (SPE) column, which integrated extraction and cleanup into one single-step.
In a typical procedure, 2.0 mL of AgNO3 solution (2.0 × 10−3 mol L−1) and 2.0 mL of KH2PO4 buffer solution (pH 7.0) were transferred into 10-mL volumetric flasks.
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CEO of Professional Science Editing for Scientists @ prosciediting.com