Author: Lilian Liu

(A1) heat induction.
1. pick 4~5 L4s and put them into new NGM plates (4~5 plates).

Preparation 6X solution

Ampicillin Stocks & Usage

* Stock Concentration 50mg/ml in H2O
* Working Concentration 50ug/ml 

We have had some luck with paralyzing worms with a BDM (2,3-butanedione monoxime) levamisole mix. I still have a lot of movement with just 10mM levamisole, but this mix is great.

While working with RNA:

Wipe down area with RNAzap.
Have dedicated “RNAse free” tips, solutions, etc.
Use filter tips.

Fosmid recombineering (Kerri Spilker 10/5/09, based on Oliver Liu 7/6/09)

Reference: Tursun et al., 2009. A Toolkit and Robust Pipeline for the Generation of Fosmid-based Reporter Genes in C. elegans. PLOS One.

For mutagenesis, one wants to prepare 6-10 plates of animals enriched for the L4 stage. Pick plates of 4 or 5 L4 animals 4 days before mutagenesis and put in 20 C incubator. In the end you only need about 4 plates for mutagenesis.

This protocol describes how to cause RNAi-induced gene silencing via feeding. Use the RNAi section of the intralab site to identify frozen bacterial strains for your gene(s) of interest.

This protocol can be used to isolate genomic DNA from one plate of worms.

1. Pick 100 L4 worms containing the array of interest into an empty plate. If you have several arrays of the same construct, use one which has a low transmission rate, as it will require less generations to isolate an integrant.