The clones are available as glycerol stocks, plates, DNA minipreps or midipreps.
If you need virus particles produced from the DNA preps, we can forward the DNA directly to the HelVi BVC virus core. Please contact them diretly to arrange for the virus production.
Clones are distributed to Biocenter-Finland affiliated groups only.
Our collection comprises 81 534 human shRNAs and 77 994 mouse shRNAs. There are usually five different constructs that target the same gene. The constructs are in pLKO.1 lentiviral vector with puromycin selection. Carbenicillin selection is used for the growth and maintenance of the vector in bacterial cells. See the vector map and sequence here.
More detailed information on the constructs can be found at the manufacturer website.
Negative controls in our stock:
SCH001: Empty pLKO.1
SHC002: Scrambled shRNA in pLKO.1
SHC016: Non-targeting shRNA in pLKO.1
Positive control:
SHC003: TurboGFP in pLKO.1
There are 40 803 mouse and 34 334 human gRNA constructs in our library. The constructs are in LV04 lentiviral vector that includes pyromycin selection cassette and a blue fluorescent protein (BFP) tag for positive-cell visualization. Ampicillin is used for selection in bacteria. The full vector map and sequence can be found here.
You can find more information on the clones on the distributor website: Sanger KO CRISPR-library.
Available negative controls:
CRISPR19: Non-target control
CRISPR20: Non-target control
To order clones or DNA, fill the order form and send it to bch-gbu@helsinki.fi.
The delivery time is approximately one week.
We always recommend to sequence the clones before the experiments. The libraries are vendor-verified but unfortunately do contain some errors. You can find the sequencing primers suitable for your clone in the info sheet we send with your order. We offer a service to verify the clones by Sanger sequencing (approximately two days) or restriction analysis (one day). Please indicate in the form if you would like to purchase also the verification service and which method.
Please indicate in the form if you would like to submit the DNA directly to the HelVi BVC for virus production. We will provide BVC with enough DNA for the desired product. Please contact bvc-support@helsinki.fi for more information and orders.
DNA preps are delivered by mail. Glycerol stocks are shipped with UPS or FedEx on dry ice on the customer's expense (customer's account number required for FedEx). For plates the delivery options are pick-up and UH internal mail.
The clones are provided free of charge to Biocenter Finland groups. A handling fee is charged, however, covering the storage, handling, and packing the clones. DNA preparations are charged for work and reagents, 0 % VAT.
UH customer | BF customer | |
---|---|---|
DNA MINIprep from the TRC or Sanger library | ||
1st item in batch | 36 € | 40 € |
additional items | 19 € | 23 € |
DNA MIDIprep (minimum 50ug) from the TRC or Sanger library Only for orders continuing to virus production |
||
1st item in batch | 60 € | 67 € |
additional items | 30 € | 37 € |
Glycerol stock (no validation) | ||
1st item in batch | 20 € | 23 € |
additional items | 15 € | 18 € |
Plate (no validation), pick-up or UH internal mail only | ||
1st item in batch | 20 € | 23 € |
additional items | 15 € | 18 € |
Clone validation by Sanger sequencing | ||
1st item in batch | 20 € | 22 € |
additional items | 15 € | 17 € |
Clone validation by restriction analysis | ||
1st item in batch | 15 € | 17 € |
additional items | 5 € | 6 € |
A batch is defined as samples of one customer processed at the same time.
Usage of clones and acknowledgements
Please indicate GBU contribution in your publications (materials and methods section/acknowledgements) e.g. in the following way:
For TRC1 shRNA clones:
These clones (xx, yy) were from Sigma Aldrich MISSION shRNA library, distributed by Genome Biology Unit core facility supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland.
For CRISPR clones:
These clones (xx, yy) were from Sanger gRNA library, distributed by Genome Biology Unit core facility supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland.
Please note that these reagents are solely for research purposes. See MTA (Material Transfer Agreement) at the Sigma web page.