Journal: microPublication Biology

Article Title: Investigation of PAL-1 requirement in C. elegans physiology using the auxin-inducible degradation system

doi: 10.17912/micropub.biology.001057

Figure Lengend Snippet: (a) Schematic of AID*::3xFLAG::wrmScarlet knock-in at the pal-1 locus (referred to as PAL-1 ::degron), and primers used to genotype for cassette insertion. Scale bar indicates 500 base pair. (b) PCR gel to illustrate wildtype (512 bp), heterozygote knock-in (512 bp and 881 bp), and homozygous knock-in (881 bp) animals . All 3 primers were used in a single PCR to differentiate the 3 possible genotypes. The P1-P3 combination producing 1406 bp is not visible in the homozygous PAL-1 ::degron strain likely due to the preferential amplification of a shorter PCR product from P1-P2. Banding patterns observed below the 500 bp are likely due to non-specific amplification or primer dimers. (c) Effects of ethanol and auxin treatment on body length of PAL-1 ::degron C. elegans ( MWU206 ) starting from L1. N=98-116 animals scored per condition. The scatter dot plot line indicates mean ± SD. (d) The Nob phenotype was observed in a small percentage (<1%) of auxin-treated MWU206 animals, the scale bar is 500 µm. Effects of ethanol and auxin treatment on (e) egg hatch rate of MWU206 (N= 7 replicates per condition with 36-91 offspring/eggs scored per replicate; scatter dot plot line indicates mean ± SD.), (f) MWU206 lifespan when fed with empty vector (EV) or ccf-1 RNAi bacteria grown on ETOH or auxin from L1; N > 125 animals were scored for each condition.

Article Snippet: PHX6872 , pal-1 ( syb6872 ) , SunyBiotech.

Techniques: Knock-In, Amplification, Plasmid Preparation, Bacteria