| Antibodies |
| Polyclonal rabbit anti-GFP primary antibody |
Novus Biologicals |
NB600-308 |
| Alexa Fluor 488-conjugated goat anti-Rabbit IgG secondary antibody |
ThermoFisher |
A11034 |
| Bacterial and virus strains |
|
Escherichia coli OP50-1 |
CGC |
N/A |
| Chemicals, peptides, and recombinant proteins |
| Protease from Streptomyces griseus |
Sigma |
P-6911 |
| Vectashield with DAPI Mounting Medium for Fluorescence |
Vector Laboratories |
H-1200 |
| S. pyogenes Cas9 (10ug/uL) |
IDT |
1081058 |
| Nuclease-free Duplex buffer |
IDT |
11- 01-03-01 |
| Invitrogen™ Wheat Germ Agglutinin, Tetramethylrhodamine Conjugate |
Fisher Scientific |
W849 |
| Critical commercial assays |
| Zymo Direct-zol RNA MicroPrep kit |
Zymo Research |
R2061 |
| LunaScript RT SuperMix kit |
NEB |
E3010S |
| Phusion high fidelity DNA polymerase |
NEB |
M0530S |
| Expand Long Template PCR system |
Roche |
11681834001 |
| NEBuilder® HiFi DNA Assembly Master Mix |
NEB |
E2621S |
| Miniprep kit |
Qiagen |
27104 |
| PCR purification kit |
Qiagen |
28104 |
| MinElute PCR purification kit |
Qiagen |
28004 |
| Gel extraction kit |
Qiagen |
28704 |
| LongAmp Taq PCR kit |
NEB |
E5200S |
| Competent cells |
NEB |
C2987H |
| KAPA HiFi HS+dNTPs (100U) |
Roche |
KK2501 |
| KAPA Hyper Prep PCR-free (8rxn) |
Roche |
KK8501 |
| AMPure XP beads |
Beckman Coulter |
A63880 |
| Experimental models: Organisms/strains |
|
C. elegans: Strain N2 |
Caenorhabditis Genetics Center (CGC) |
N2 |
|
C. elegans: Strain BA829 spe-36(it114) unc-22
|
Ken Kemphues, Cornell University |
BA829 |
|
C. elegans: Strain AD99 spe-36(as1)
|
This study |
AD99 |
|
C. elegans: Strain AD105 spe-36(as6)
|
This study |
AD105 |
|
C. elegans: Strain CB4856 Hawaiian mapping strain |
CGC |
CB4856 |
|
C. elegans: Strain DR466 him-5(e1490)
|
CGC |
DR466 |
|
C. elegans: Strain BA17 fem-1(hc17)
|
CGC |
BA17 |
|
C. elegans: Strain CB61 dpy-5(e61)
|
CGC |
CB61 |
|
C. elegans: Strain AD348 glp-4(bn2); him-5(e1490)
|
Cross between SS104 and DR466 |
AD348 |
|
C. elegans: Strain MDX44 cylc-2(mon2[cylc-2::mNG^3xFLAG])
|
Krauchunas et al., 202021
|
MDX44 |
|
C. elegans: Strain SL438 spe-9(eb19) I; him-5(e1490) V; ebEx126
|
CGC |
SL438 |
|
C. elegans: Strain RT36 arIs37 [myo-3p::ssGFP]
|
Barth Grant, Rutgers University |
RT36 |
|
C. elegans: Strain HCL67 unc-119(ed3) III; uocIsl [eft-3p::Cas9(dpiRNA)::tbb-2 3′ UTR + unc-119(+)]
|
CGC59
|
HCL67 |
|
C. elegans: Strain ARK5 spe-36(as6); asEx96 [PCR product of genomic spe-36]
|
This study |
ARK5 |
|
C. elegans: Strain ARK6 spe-36(as6); him-5(e1490); asEx96 |
This study |
ARK6 |
|
C. elegans: Strain ARK7 spe-36(nwk1[spe-36::gfp]); him-5(e1490) |
This study |
ARK7 |
|
C. elegans: Strain ARK17 cylc-2(mon2); spe-36(as6); him-5(e1490); asEx96
|
This study |
ARK17 |
|
C. elegans: Strain AD349 asEx112 [myo-3p::spe-36::gfp]
|
This study |
AD349 |
|
C. elegans: Strain AD350 asEx113 [myo-3p::spe-9::gfp]
|
This study |
AD350 |
| Oligonucleotides |
|
|
| PCR Primer for TruSeq library amplification, SG – 915 AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGA |
Sam Gu, Rutgers University, designed by Illumina |
n/a |
| PCR Primer for TruSeq library amplification, SG – 916 CAAGCAGAAGACGGCATACGAGAT |
Sam Gu, Rutgers University designed by Illumina |
n/a |
| PCR Primer to amplify F40F11.4 plus 479 bp upstream and 303 bp downstream GGAAGAATGTAACGGCTGCTATC |
This study |
n/a |
| PCR Primer to amplify F40F11.4 plus 479 bp upstream and 303 bp downstream CCGGATTGAGGTTGGTGTTT |
This study |
n/a |
| PCR Primer to amplify spe-36 for Gibson assembly CCCACGACCACTAGATCCATCTAGAAAAAAATGAATTTCAAAATATGTATTCTGTTC |
This study |
n/a |
| PCR Primer to amplify spe-36 for Gibson assembly TCCTTTACTCATTTTTTCTACCGGTAACAATTCATCCATCTCTTCAG |
This study |
n/a |
| PCR Primer to amplify spe-9 for Gibson assembly CCCACGACCACTAGATCCATCTAGAAAAAAATGAATGTGATTCTGGTG |
This study |
n/a |
| PCR Primer to amplify spe-9 for Gibson assembly TCCTTTACTCATTTTTTCTACCGGTAAAAATGGATTCTCAGTTTTCAC |
This study |
n/a |
| PCR primers for RT-PCR can be found in Table S1
|
|
|
| Oligos and RNAs for CRISPR can be found in Table S2
|
|
|
| Recombinant DNA |
| PJF25 |
Barth Grant, Rutgers University57
|
|
|
myo-3p::spe-36::gfp (signal peptide replaced with spe-36 cDNA in PJF25) |
This study |
|
|
myo-3p::spe-9::gfp (signal peptide replaced with spe-9 cDNA in PJF25) |
This study |
|
| Software and algorithms |
| MimodD |
https://mimodd.readthedocs.io/en/latest/
|
|
| FUSION |
https://fusion.help.andor.com/display/fusionum/Introduction
|
|
| Imaris |
https://imaris.oxinst.com/
|
|
| Zeiss Zen |
https://www.zeiss.com/microscopy/en/products/software/zeiss-zen.html
|
|
| GraphPad Prism |
https://www.graphpad.com/
|
|
