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.
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. Because we can't guarantee the quality of customer's own plasmid we can't guarantee the success of the mutagenesis. We will make two trials but will need to charge them independent of the success. Failed mutagenesis is rare (<10%).
The prices have been updated and these prices are effective from March 1st 2024.
| Customer class | University of Helsinki, First item |
University of Helsinki, subsequent items |
Biocenter Finland, first item |
Biocenter Finland, subsequent items |
External, fisrt item |
External, subsequent items |
|---|---|---|---|---|---|---|
| BP-cloning (PCR product to entry vector) | 80 € | 25 € | 93 € | 30 € | 330 € | 56 € |
| LR-cloning (entry clone to expression vector) | 65 € | 30 € | 77 € | 34 € | 293 € | 60 € |
| Mutagenesis | 150 € | 75 € | 173 € | 89 € | 584 € | 224 € |
| Control construct | 20 € | 20 € | 20 € | 20 € | 40 € | 40 € |
Examples:
Cloning of an ORFeome clone to an expression vector: clone miniprep 36 € + LR cloning to an expression vector: 65 €, total 101 €, second clone+cloning 19 € + 30 € = 49 €.
Cloning of your own PCR product to expression vector: PCR primer design free of charge, BP cloning to entry-vector: 80 €, LR cloning to an expression vector: 65 €, total 145 €, second 25 € + 30 € = 55 €.
Mutagenesis of library clone: Miniprep from library: 36 €, mutagenesis: 150 €, LR-cloning to expression vector: 65 €, total 251 €.
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
Haartmaninkatu 8
P.O.Box. 63
00014 University of Helsinki
We collaborate with Biomedicum Virus Core (BVC) and can deliver your lenti-construct there directly. Please contact bvc-support@helsinki.fi to agree on the virus production.
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"
ORFeome clones:
“Name of the clone (ORFeome Library; Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland)"
MGC clones:
“Name of the clone (MGC Library; Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland)"