| Antibodies |
| His Tag biotinylated antibody | R&D Systems | BAM050 |
| Biological samples | | |
| Purified tubulin from bovine brains | Schenk Packing Co. | NA |
| Yeast whole-cell lysate | Listed strains | NA |
| Chemicals, peptides, and recombinant proteins |
| Tubulin protein (fluorescent HiLyte 647): Porcine brain | Cytoskeleton, Inc. | TL670M |
| Tubulin protein (biotin): Porcine brain | Cytoskeleton, Inc, | T333P |
| Vectabond reagent, tissue section adhesion | Vector Laboratories | SP-1800-7 |
| mPEG-SVA MW 5,000 | Laysan Bio, Inc. | mPEG-succinimidyl valerate |
| Biotinylated mPEG-SVA MW 5,000 | Laysan Bio, Inc. | Biotin-PEG-SVA |
| Avidin DN | Vector Laboratories | A-3100-1 |
| Biotin-BSA | Vector Laboratories | B-2007-10 |
| Paclitaxel | Millipore Sigma | T7191-5MG |
| Guanosine 5′-triphosphate sodium salt hydrate | Milipore Sigma | G8877-1G |
| Streptavidin-coated polystyrene beads | Spherotech | SVP-05-10 |
| Guanosine-5′-[(α,β)-methyleno]triphosphate, sodium salt (GMPcPP) | Jena Bioscience | JBS-NU-405S |
| Experimental models: Organisms/strains |
| S. cerevisiae: MATa ura3-1::pCSE4-CSE4-XbaI(GFP):URA3 leu2,3-112 his3-11 trp1-1 ade2-1 LYS2+ can1-100 bar1 cse4Δ::KanMX | Biggins lab | SBY19926 |
| S. cerevisiae: MATa pDsn1-Dsn1-2D-3FLAG:URA3 Ndc10-GFP:kanMX6 | Biggins lab | SBY21618 |
| S. cerevisiae: MATa pDsn1-Dsn1-2D-3FLAG:URA3 Ndc80-GFP:kanMX6 | Biggins lab | SBY21620 |
| S. cerevisiae: MATa pDsn1-Dsn1-2D-3FLAG:URA3 Ctf19-GFP:kanMX6 | Biggins lab | SBY21621 |
| S. cerevisiae: MATx pDsn1-Dsn1-2D-3FLAG:URA3 Dam1-3GFP:HIS | Biggins lab | SBY20634 |
| Oligonucleotides |
| Forward primer for generating 208-bp CEN3 or CEN3mut: /5BiosG/5′-GGTGGTTCTGGTGGTTCTGGTGAATTC-CCATTCAATGAAATATATATTTCTTACTATTTC-3′ | This paper | 50_JDL |
| Reverse primer for generating 208-bp CEN3 or CEN3mut: /5Atto565N/5′-GCTATTCATTGAAAAAATAGTACAAATAAG-3′ | This paper | 52_JDL |
| 5′ primer to tag Ndc80: 5′-ACGAAATTTGGAGTTTGAAACTGAACATAACGTAACAAATCGGATCCCCGGGTTAATTAA-3′ | This paper | 30_JDL |
| 3′ primer to tag Ndc80: 5′-CTGTAGATTGCTCGGGTATTATATATCATTTATTTTATTAGAATTCGAGCTCGTTTAAAC-3′ | This paper | 31_JDL |
| 5′ primer to tag Ctf19: 5′-GATCTGCAACGTTTGCCTATTCCCGGACATGTACGCCAGGCGGATCCCCGGGTTAATTAA-3′ | This paper | 10_JDL |
| 3′ primer to tag Ctf19: 5′-TAAGCAAGCCGTCCAGTTGGCAATGGCAAATGGAACATCAGAATTCGAGCTCGTTTAAAC-3′ | This paper | 11_JDL |
| 5′ primer to tag Ndc10: 5′-TCAAAATTCATTTGATGGTCTGTTAGTATATCTATCTAACCGGATCCCCGGGTTAATTAA-3′ | This paper | 6_JDL |
| 3′ primer to tag Ndc10: 5′-TATCCCTATACGAAACAGTTTAAACTTCGAAGCTCCCTCAGAATTCGAGCTCGTTTAAAC-3′ | This paper | 7_JDL |
| Recombinant DNA |
| pSB963: WT CEN3, 8LacO, TRP1 | Lang et al. (2018) | NA |
| pSB972: Mutant CEN3 (CCG->AGC CEN mutant), 8 LacO, TRP1 | Lang et al. (2018) | NA |
| Software and algorithms |
| Single molecule colocalization analysis (written in LabView) | This paper | https://github.com/casbury69/smTIRF-spot-selection-colocalization-and-brightness-vs-time |
| Displacement analysis (written in Igor Pro) | This paper | https://github.com/casbury69/flip-flop-IGOR-analysis-routines |
| ImageJ (Fiji) | Schindelin et al. (2012) | https://imagej.net/software/fiji/ |
| MOSAICsuite | Sbalzarini and Koumoutsakos (2005) | https://imagej.net/plugins/mosaicsuite |