Cloning into expression vectors starting from cDNA or genomic DNA :

  PCR amplification of the requested constructs, restriction cloning or recombination in bacterial vectors ;
  mini-preparation of the vectors (Macherey-Nagel) ;
  constructs will be sequenced upon request

Constructions
  Vectors: derived from pET (NOVAGEN) ; pET9 - pET28 - pET21 (restriction sites 5’: NdeI, NcoI or NheI respectively). Cloning in NotI site for 3’.
  Gateway system (upon special request)
  Tagging: His-Tag on C-terminal or N-terminal. Two options:

• Tag included in 3’ primer.
• Tag after restriction (introducing a 3 amino acid linker).

Delivery: all the cloning will be done on the MWG cloning robot and for optimal use experiments will be grouped into 96 well plates; delivery can therefore be variable but will be discussed beforehand with the customer

Price:

1. cloning with primers and sequencing: 300€/construction
2. cloning with only primers: 225€/construction
3. cloning with only sequencing: 150€/construction
4. cloning only: 60€/construction

Contact: sophie.cheruel@egm.u-psud.fr