|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||15182||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backboneRCAS (A)
- Backbone size w/o insert (bp) 12000
Vector typeRetroviral, RNAi ; Avian expression
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth Strain(s)ccdB Survival
Growth instructionsGrow in ccdB Survival or DB3.1 cells
Gene/Insert nameGateway cassette
- Cloning method Restriction Enzyme
- 5′ sequencing primer n/a (Common Sequencing Primers)
Terms and Licenses
The RCAS-RNAi viruses have been modified so that they are now compatible with the Gateway cloning system (Invitrogen). DNA encoding the hairpin is cloned into a modified U6 vector using standard cloning techniques. The construct is then moved into the retroviral vector using Gateway reagents, thus bypassing the difficulties of cloning small fragments into RCAS. As a result, each construct can be made with a minimal number of cloning steps. See Supplemental Information of the article for a detailed protocol.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:G-RCAS (A) was a gift from Connie Cepko (Addgene plasmid # 15182 ; http://n2t.net/addgene:15182 ; RRID:Addgene_15182)
For your References section:RCAS-RNAi: a loss-of-function method for the developing chick retina. Harpavat S, Cepko CL. BMC Dev Biol. 2006 . 6():2. 10.1186/1471-213X-6-2 PubMed 16426460