Site specific mutagenesis is; a technique used to:
- create specific alterations in a gene
- study the relationship between protein structure and its function
- modify or repair DNA constructs
- alter sites of restriction, insert or delete elements of a vector (for example tag elements, or exchange of promoters)
- We can introduce any type of single or multiple mutation * (substitution, insertion or deletion) in plasmids up to 15kb.
We do not recommend the use of more plasmids; large (> 15 kb, lentiviral ...), low copy, or with very repeated sequences or rich in GC because they could easily rearrange or be inefficient in replication. In these cases, contact the laboratory by writing to This email address is being protected from spambots. You need JavaScript enabled to view it.

THE SERVICE INCLUDES:
- oligo-sense sequencing of the portion to be mutagenized to confirm DNA before mutagenesis
- design and synthesis of the mutagenic oligo
- DNA amplification
- transformation, cell culture and purification of 5 random colonies
- sequencing of purified clones to confirm mutation
- 4/5 working days for single mutation
- sending of alignment with the wt control sequence and single electropherogram of the positive clone
- dispatch of 1/4 µ g of purified mutated plasmid DNA

ON REQUEST:
- sequencing of the entire insert or plasmid with synthesis of the relative oligos that will be stored in the personal Oligo Bank
- control of the plasmid by digestion with restriction enzymes
- major preparation of purified plasmid DNA (midi or maxi)
- sequencing of the site to be mutagenized and of the positive clone also with anti-sense oligo.

It should be noted that the completion of all quality controls provided, while not guaranteeing the functioning of the clone, will in any case give Bio-Fab the right to receive the agreed payment up to the stage of the project carried out.

MATERIAL TO BE SHIPPED TO THE LABORATORY:
- 2 µg of purified plasmid containing the DNA to be mutated, from a strain of E. coli dam +. Plasmids isolated from dam- strains are not suitable (e.g. jm110, scs110 and BL21)
- plasmid map with indication of antibiotic resistance and target DNA sequence
- mutagenesis instructions: list and localization of the bases for insertion, deletion and replacement
- form completed in its entirety

*for the cost of multiple mutations please contact This email address is being protected from spambots. You need JavaScript enabled to view it.

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