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Analysis of gene expression between monomethyl branched-chain faatty acid deficient (elo-5) and wild type N2 C. elegans L1 larvae hatched on food-free NMG plates

ABSTRACT: Gene expresssion is not globally shut down in elo-5 as compared to N2 Many changes in metabolic and regulatory gene expression were detected between elo-5 and N2 L1 larvae. RNA samples for hybridization with the C. elegans Affymetrix Gene Chips were prepared as follows. elo-5(gk208) and N2 control animals were grown from eggs for one generation on NGM plates spotted with 300Âµl OP50 overnight liquid culture supplemented with 1mM C17ISO in DMSO at 20Â°C. Gravid adults were washed off the plates with M9 and bleached. Eggs were plated on OP50/NGM plates supplemented with 1mM of either C13ISO or C17ISO and maintained at 20Â°C to obtain gravid adults. Adults were washed off the plates and bleached, and eggs were then plated on food-free NGM or OP50/NGM plates at 20Â°C. Twenty-four hours later, L1s from food-free plates were collected for RNA isolation with Trizol and Qiagenâ??s RNAse Easy Kit, according to the manufacturersâ?? protocols. On the following days, animals on OP50/NGM were checked for phenotypes to confirm C17ISO-deficiency in elo-5(gk208) grown with C13ISO supplement. Two biological replicates were prepared for each condition; elo-5(gk208) with C13ISO and C17ISO supplements and a control N2 with C13ISO supplement. Data analysis was performed with dChip analytical software (Li and Wong, 2001).

ORGANISM(S): Caenorhabditis elegans

SUBMITTER: Marina Kniazeva

PROVIDER: E-GEOD-64973 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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