
                                      PROCEDURE
Prothrombin time is typically analyzed by a laboratory technologist on an automated instrument at 37 °C (as a nominal approximation of normal human body temperature).
Blood is drawn into a test tube containing liquid sodium citrate, which acts as an anticoagulant by binding the calcium in a sample.
 The blood is mixed, then centrifuged to separate blood cells from plasma (as prothrombin time is most commonly measured using blood plasma). In newborns, a capillary whole blood specimen is used.
A sample of the plasma is extracted from the test tube and placed into a measuring test tube (Note: for an accurate measurement, the ratio of blood to citrate needs to be fixed and should be labeled on the side of the measuring test tube by the manufacturing company; many laboratories will not perform the assay if the tube is underfilled and contains a relatively high concentration of citrate—the standardized dilution of 1 part anticoagulant to 9 parts whole blood is no longer valid).
Next an excess of calcium (in a phospholipid suspension) is added to the test tube, thereby reversing the effects of citrate and enabling the blood to clot again.
Finally, in order to activate the extrinsic / tissue factor clotting cascade pathway, tissue factor (also known as factor III) is added and the time the sample takes to clot is measured optically using calorimeter or photometer.