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GFP::PCN-1 does not reliably mark S phase in C. elegans adult germline progenitor zone cells

Tokiko Furuta & Swathi Arur
PCNA (proliferating cell nuclear antigen) is the DNA polymerase processivity factor that loads onto the chromatin during S phase of the cell-cycle (Brauchle et al. 2003). Thus, nuclear localization of PCNA (PCN-1 in C. elegans) is used as a marker for the S phase of the cell cycle (Brauchle et al. 2003). GFP::PCN-1 has been shown to label S phase in C. elegans embryo when driven through the germline and embryonic promoter pie-1 (Brauchle et...

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Edwin Soedarmadji, John Gregoire & Contributors

Establishment of a CRISPR/Cas9-based strategy for inducible protein dimerization

Jeffrey Zielich, Sriyash Mangal, Esther Zanin & Eric J. Lambie
We modified Norris’s mCherry-tag repair donor vector by fusing a C. elegans codon-optimized fkbp12 sequence 3’ to mCherry (Mangal et al. 2018) and re-establishing the critical NotI site for 3’ homology region insertion (Figure 1 A; pEL226). This vector can be used to C-terminally tag any GOI by following the protocol of Norris et al. (2015), including subsequent Cre-mediated excision of the dual marker selection cassette (Figure 1 B). As proof of principle we C-terminally...

oxi-1 and fshr-1 are required for neuromuscular signaling under normal and oxidative stress conditions in C. elegans

Barry Wei & Jennifer R Kowalski
Reactive oxygen species (ROS) contribute to neuronal degeneration by readily reacting with cellular components, consequently breaking down cellular integrity. Excess ROS often leads to oxidative stress, which results from destabilization of the organism's ability to control the balance between antioxidants and free radicals (Chandra et al. 2015). The ubiquitin-proteasome system helps to regulate oxidative stress and overall damage to cellular components by forming chains of ubiquitin polypeptides on cellular proteins; these chains then serve as...

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Edwin Soedarmadji, John Gregoire & Contributors

Simplified detection of a point mutation in C. elegans using tetra-primer ARMS-PCR

Matthew T Sullenberger & Eleanor M Maine
Single nucleotide polymorphisms (SNPs) can be difficult to detect using traditional PCR and gel electrophoresis, especially when no enzyme restriction sites overlap the SNP. Although alleles can be identified through amplification with flanking primers and sequencing, this adds time and cost, and sequencing is typically outsourced. Tetra-primer Amplification-Refractory Mutation System (ARMS)-PCR allows SNPs to be distinguished through a 3' primer mismatch at the SNP site for one allele, and an additional mismatch 1 - 3...

Materials Experiment and Analysis Database:

Edwin Soedarmadji, John Gregoire & Contributors

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