dsRNA was produced in vitro using the clones from the Ahringer feeding library (Fraser et al., 2000 (link); Kamath et al., 2003 (link)). The clones used in this study are listed in Table S2. For the control, we used the clone C06A6.2 previously found in the laboratory to have no effect on the early embryonic cell division and to be 100% viable. Each template was amplified by PCR using T7 primers, and dsRNA was produced using the Promega Ribomax RNA production system. dsRNA was injected in L4/young adult hermaphrodites. The incubation time of the injected worms before embryo dissection is listed in Table S2.
Ribomax rna production system
Manufactured by Promega
The Ribomax RNA production system is a laboratory equipment designed for the in vitro synthesis of RNA. It provides the necessary components and protocols to generate high-quality RNA samples for various research applications.
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Lab products found in correlation
2 protocols using ribomax rna production system
1
In Vitro dsRNA Production for RNAi
2
Genome-wide RNAi screen in C. elegans
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A list of the genes silenced through RNAi in this study is provided in Table S4 .
Clones from the Ahringer feeding library (Ahringer, 2006 (link); Kamath et al., 2003 (link)) were used when available. As a control, we used the clone C06A6.2, previously found in the laboratory to not affect early embryonic division and development (injected worms are 100% viable). To produce pqn-59 dsRNA, a DNA fragment was amplified from genomic DNA using Gateway-compatible oligonucleotide primers (as inTable S4 ) for Gateway-based-cloning into the pDESTL4440 plasmid (Addgene plasmid #11344 ). The DNA was subsequently amplified using standard T7 primers. For tiar-1, the DNA was amplified from genomic DNA using oligonucleotides with T7 overhangs (see Table S4 ). For all genes, the dsRNA was produced with the Promega Ribomax RNA production system. dsRNA was injected in L4 or young adult hermaphrodites, which were incubated at 20°C. Germlines or embryos collected from injected hermaphrodites were analyzed 24 h after injection.
Clones from the Ahringer feeding library (Ahringer, 2006 (link); Kamath et al., 2003 (link)) were used when available. As a control, we used the clone C06A6.2, previously found in the laboratory to not affect early embryonic division and development (injected worms are 100% viable). To produce pqn-59 dsRNA, a DNA fragment was amplified from genomic DNA using Gateway-compatible oligonucleotide primers (as in
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