Design and order your oligos according to these instructions by Addgene. Note that the ends of the oligos must be exactly as detailed in the protocol for the cloning to work. We can verify your oligo sequences if needed. Deliver the oligos to us and we can proceed with the cloning.
Oligonucleotide ordering tips: Standard de-salted oligos (usually the most inexpensive synthesis) are sufficient for cloning. If not already resuspended, dilute each oligo to 100 μM in DNAse-free water.
The shRNA oligos are annealed and cloned to TRC library vector pLKO.1 (Moffat et al Cell. 2006 Mar 24. 124(6):1283-98), a lentivector with puromycin resistance for selection in mammalian cells. The clones are verified with Sanger sequencing, and the customer will receive a DNA miniprep of each of the the clones. Bacterial glycerol stocks will also be prepared and can be picked up from GBU if needed.
The clones can be submitted directly to HelVi BVC for virus production. Please contact bvc-support@helsinki.fi for more information and orders.
| Customer class | First cloning in order | ADDitional reactions |
|---|---|---|
| University of Helsinki | 100 € | 60 € |
| Biocenter Finland | 115 € | 70 € |
| External | 475 € | 165 € |
If you use a clone from the GBU in your publication, please acknowledge us in the following way:
“shRNA constructs were generated by the Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland"
Please also cite the creators of the vector backbone of your construct: pLKO.1 (Moffat et al Cell. 2006 Mar 24. 124(6):1283-98)