G-RCAS (A)
(Plasmid #15182)

• Depositing Lab: Connie Cepko
• Publication: Harpavat et al BMC Dev Biol. 2006 . 6():2.

Backbone

• Vector backbone: RCAS (A)
• Backbone size w/o insert (bp): 12000
• Vector type: Retroviral, RNAi ; Avian expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Growth instructions: Grow in ccdB Survival or DB3.1 cells
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Gateway cassette

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: n/a

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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.