GBU performs Gateway cloning to various expression vectors, and site-directed mutagenesis of our ORF library clones.
We do cloning from the ORFeome collection, as well as from your own Gateway-compatible PCR products and plasmids to a number of destination vectors. Our selection of vectors includes a wide range of expression vectors ranging from bacterial and vertebrate expression to lentivectors. If your favourite vector is not on our list, feel free to suggest an update to our collection.
Additionally we provide help in designing primers to create Gateway-compatible PCR products from non-Gateway-compatible templates. We can then clone the PCR products to destination vectors in two cloning steps.
The cloned products are delivered as transfection grade minipreps and glycerol stocks. Minipreps can be delivered by mail if necessary.
You can make your cloning order with the same form as the library clone orders.
The following control constructs can be cloned to all destination vectors.
- EmGFP with stop
- EmGFP without stop
- mCherry with stop
- mCherry without stop
- lacZ with stop
- lacZ without stop
- empty insert (the destination vectors with this insert contain no ccdb gene and can be grown in DH5alpha and stbl3 competent cells)
With our site-directed mutagenesis service, we can generate one to a few nucleotide sequence alterations to our library-derived plasmids.
The mutagenesis service fee covers the site-directed mutagenesis reaction (two tries), as well as two miniprep DNA extractions and sequencing of two mutated plasmid clones. An additional fee is charged when competent cells than standard Dh5alpha are needed (e.g. for lentiviral plasmids).
We use a Phusion HF polymerase with an error rate of 4,4 x 10-7, and while random mutations caused by the polymerase are rare, they are still possible. We confirm the mutated site by sequencing, but we do not guarantee that the construct is free of replication errors outside the mutated region.
We primarily do mutagenesis service only for the clones derived from our libraries. If you would like to have your own construct mutated, please contact us, and we will evaluate each service request case-by-case. The map and the full vector sequence are needed for the approved services. We will try the mutagenesis twice and in case of customer's own plasmid will charge the reaction anyway even if we don't get a successful clone.
|Customer class||University of Helsinki||Biocenter Finland||External|
|Cloning reaction||40 €||40 €||80 €|
first item in order
|180 €||190 €||380 €|
(max. batch 8 reactions)
|90 €||95 €||180 €|
|Control construct||20 €||20 €||40 €|
Cloning of an ORFeome clone to an expression vector: processing fee of the clone: 20€, LR cloning to an expression vector: 40€, total 60€.
Cloning of your own PCR product to expression vector: PCR primer design free of charge, BP cloning to entry-vector: 40€, LR cloning to an expression vector: 40€, total 80€.
GBU also performs cloning reactions to customer-provided Gateway-compatible entry-constructs and PCR products. Please, deliver your samples to GBU sample fridge (Biomedicum Helsinki 1, 5th floor, room B505b). If you send samples by mail please use the address below:
Genome Biology Unit , 5th floor, rooms B505b ja B502a
Biomedicum Helsinki 1
00014 University of Helsinki
If you use a clone from the GBU in your publication, please acknowledge us in the following way:
Cloning and mutagenesis:
“Constructs were generated by the Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland"
“Name of the clone (ORFeome Library; Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland)"
“Name of the clone (MGC Library; Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland)"