Isolation DNA using AS WHOLE BLOOD SAMPLES (1 small yellow top).
a. Centrifuge small yellow top tubes for 10 minutes at 2500RPM. Discard top part of the serum and mixed the rest of the blood three times by inverting to mix the buffy coat with the the rest of the red cells.
b. Per ASHI sample: transfer 350µl of the blood mixture into three 2.0ml sterile sample tubes.
c. Insert the EZ1® Whole Blood Card into the card slot. Turn on the EZ1® BioRobot and open the workstation door.
d. Invert each reagent cartridge twice to mix the magnetic particles. Then tap the reagent cartridges to deposit the reagents at the bottom of their wells. Load the EZ1® cartridge rack with three reagent cartridges per ASHI sample (See procedural notes).
e. Load the EZ1® worktable: the uncapped 1.5 ml elution tubes in the 1st row, the disposable filter tips with disposable tip holders in row 2 (row 3 is empty), and the ASHI sample uncapped 2.0ml sample tubes with 350µl whole blood in the 4th row of the worktable. Double check this order by following the protocol instructions listed on the LCD screen of the instrument. (See procedural notes) Also, when loading the samples onto the instrument, be sure to double check and make sure the sample tube and elution tube labels match for each patient and are lined up on the machine correctly.
f. Select protocol #2 (350µl whole blood). Press enter.
g. Choose 200µl as the final elution volume. Choose NO to pure ethanol wash.
h. Proceed to the direction on the screen to the “Start” button and start.
DNA yield is about 50µg from 2ml of whole blood.