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Components of GMPCPP microtubule seeds, including volume and order of addition. Prepare 5 μL aliquots and store for up to 1 year at −80°C.
| Reagent | BRIGHT mix (μL) | Order of addition | BRIGHT mix + biotin (μL) | Order of addition |
|---|---|---|---|---|
| Fluorescent tubulin, 10 mg/mL | 2 | 6 | 2 | 7 |
| Biotin-tubulin, 10 mg/mL | 0 | N/A | 2 | 6 |
| Unlabeled tubulin, 10 mg/mL | 20 | 5 | 18 | 5 |
| GMPCPP, 10 mM | 30 | 4 | 30 | 4 |
| DTT, 0.2 M | 0.7 | 3 | 0.7 | 3 |
| 5X BRB80 | 26.4 | 2 | 26.4 | 2 |
| 18.2 MΩ*cm water, filtered | 52.9 | 1 | 52.9 | 1 |
| Total Volume (μL) | 132 | 132 |
