SEMA3A (chick) pBLUE
(Plasmid #29564)

• Depositing Lab: Jonathan Raper
• Publication: Raper Lab Plasmids (unpublished)

Backbone

• Vector backbone: pBLUESCRIPT (SK)
• Backbone manufacturer: stratagene
• Backbone size w/o insert (bp): 3000
• Vector type: pBlue for ISH

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SEMA3A (chick)
• Alt name: Collapsin 1
• Alt name: Semaphorin 3A
• Species: G. gallus (chicken)
• Insert Size (bp): 2100
• Mutation: 2.1kb fragment for in situ hybridization (ISH)
• GenBank ID: NM_204977
• Entrez Gene: SEMA3A (a.k.a. COLL-1, semaphorin-3A)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

To generate the sense probe for ISH, linearize the plasmid with XbaI and transcribe with T3 RNA polymerase. To generate the antisense probe, linearize the plasmid with NotI and transcribe with T7 RNA polymerase.

Please note that T7 is the 5' sequencing primer for the vector; however, the SEMA3A insert is cloned in reverse orientation. For cloning purposes, NotI is at the 5' end of SEMA3A while XbaI is at the 3' end of SEMA3A.