| Wash Step | Instructions |
| Wash #1 | Vacuum down supernatant as much as possible. This will contain oligo that did not bind to beads and sub-product, as indicated by high A260 value (This is the case for Washes #1 - #3). |
| Wash #2 | Wash with 30 mL of Quench Buffer |
| Wash #3 | Wash with 30 mL of Quench Buffer |
| Wash #4 | Wash with 30 mL of Quench Buffer |
| Wash #5 | Resuspend beads in 30 mL of Quench Buffer and incubate at room temperature for 4 hours before vacuuming off the supernatant for analysis. During and after this step sub-product will be formed, as can be seen from a high A260/A230 ratio (above 3.0). |
| Wash #6 | Wash with 30 mL of Quench Buffer |
| Wash #7 | Wash with 30 mL of Quench Buffer |
| Wash #8 | Wash with 30 mL of Quench Buffer |
| Wash #9 | Wash twice with 15 mL of Wash Buffer 2 |
| Wash #10 | Wash twice with 15 mL of Wash Buffer 2 |
| Wash #11 | Wash twice with 15 mL of Quench Buffer |
| Wash #12 | Wash twice with 15 mL of Quench Buffer |
| Wash #13 | Wash twice with 15 mL of Wash Buffer 2 |
| Wash #14 | Wash twice with 15 mL of Wash Buffer 2 |
| Wash #15 | Wash twice with 15 mL of Quench Buffer |
| Wash #16 | Wash twice with 15 mL of Quench Buffer |
| Wash #17 | Wash twice with 15 mL of Wash Buffer 2 |
| Wash #18 | Add 15 mL of Wash Buffer 2 to the beads and incubate overnight at 4 C. Always carry out this step as more sub-product will form. The next day vacuum off the supernatant and wash again with 15 mL of Wash Buffer 2. |
| Wash #19 | Wash twice with 15 mL of Quench Buffer |
| Wash #20 - X | Wash twice with 15 mL of Quench Buffer until A260 value is minimal |