C. elegans ORF-RNAi library | Source BioScience


Please note that the clone(s) being sent have tested negative in our T1 phage contamination assay. However these clones should still be handled with care as no phage assay can be guaranteed to be 100% reliable.

THE DISPATCH NOTE SHOWS THE ORIGINAL CLONE ID ORDERED AND ALSO THE ALIAS (PLATE AND WELL NUMBER). THIS IS WHAT IS SHOWN ON THE CLONE LABEL. PLEASE KEEP THE DISPATCH NOTE FOR FUTURE REFERENCE.

 

What to do with your clones when you receive them

Plates contain LB broth with 8% glycerol and ampicillin. The plates are sent on Dry Ice, and should be stored at -70 °C

Individual clones pools have been streaked onto LB agar containing ampicillin (100 µg/ml). Please store them at 4 °C (not in a freezer). As soon as possible, transfer the culture to LB broth containing ampicillin + 8% glycerol and incubate overnight at 37 °C (do not try to transfer single colonies, instead scoop out the whole bacterial culture using a disposable inoculation loop). Incubate overnight at 37 °C for subsequent freezing at -70 °C.

These clones are for research purposes only.

 

The HT115 (DE3) genotype is as follows

F-, mcrA, mcrB, IN(rrnD-rrnE)1, lambda -, rnc14::Tn10(DE3 lysogen: lavUV5 promoter -T7 polymerase) (IPTG-inducible T7 polymerase) (RNase III minus).

The universal primers that can be used for PCR amplification and sequencing are:

pL4440-dest-RNAi-FOR (5'-GTTTTCCCAGTCACGACGTT-3')
pL4440-dest-RNAi-REV (5'-TGGATAACCGTATTACCGCC-3')

Databases

Please click on ORF-RNAi.xls to view database.

 
Acknowledgements and subsequent publication

We are grateful to all the members of Marc Vidal's lab (Dana-Farber Cancer Institute) for the gift of this library and associated database.


Please acknowledge the Dana-Farber Cancer Institute and Source BioScience LifeSciences in any resulting publications.



Please also cite the following paper:


Rual JF, Ceron J, Koreth J, Hao T, Nicot AS, Hirozane-Kishikawa T, Vandenhaute J, Orkin SH, Hill DE, van den Heuvel S, Vidal M. "Toward improving Caenorhabditis elegans phenome mapping with an ORFeome-based RNAi library". Genome Res. 2004 Oct;14(10B):2162-8.
Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.