QUANTITATIVE PROCEDURE
If it is intended to routinely quantitate positive results the screening procedure may be modified by omitting the Control Cells and preparing only one final dilution. Most samples will be negative or genuinely positive, and the Control Cells may be used in the quantitative procedure below.
1. Prepare dilutions in a microtitration plate as follows:
For each sample, dispense 25l of diluent into each well in one column of the plate. For titrations of controls dispensing should commence from row 3.
Transfer 25l from row 2 of the original screening plate to row 1 of the quantitative plate.
Mix and discard 25l.
Transfer 25l from row 2 of the original screening plate to row 2 of the quantitative plate.
Prepare 25l doubling dilutions from row 2 to row 8 ( for controls doubling dilutions should commence from row 3 ).
2. Add 75l of well mixed Control Cells to row 1.
3. Add 75l of well mixed Test Cells to rows 2 to 8.
4. Mix by gentle tapping.
The final dilutions in row 1 and row 2 are 1/80.
5. Cover and let stand for 45 to 60 minutes at room temperature (or overnight).
Note: Kit controls are prediluted and 25l should be added directly into individual wells in rows 1, 2 and 3 with doubling dilutions commencing from row 3 (no diluent is required in row 1 or row 2).