Maintain tubing and holder below the site, and positioned so that the tubes fill from the bottom up to prevent reflux. Fill additive tubes until the vacuum is exhausted to ensure the correct blood-to-additive ratio and mix them immediately upon removal from the holder, using 3 to 8 gentle inversions (depending on type and manufacturer) to prevent clot formation. Follow the If a coagulation tube is the first or only tube collected, draw a discard tube first to remove air in the tubing and assure proper filling of the coagulation tube