What is the difference between MGC and ORFeome libraries?
- In ORFeome library all clones are ORFs in gateway compatible entry-vectors (pENTR221, pENTR201, pENTR223)
- In MGC library the clones are cDNAs in different vector backbones. Check the vectors from the library excel.
For more information please see:
How do I know whether you have the clone I'm interested in?
- Check the excels from the Cloning page.
What if I can't find my clone of interest?
- You can check the websites of commercial providers e.g. invitrogen, addgene, genecopoeia (check their validation status for the clones).
- If the clone you are looking for cannot be found in the excel sheets, it is unfortunately not included in our collections. However, make sure you search with all possible gene names and acronyms and please note that there are separate sheets for human and mouse constructs.
- If your clone of interest can only be found in the MGC collection but the vector is not suitable for your experiment, it is possible to produce a Gateway entry clone by PCR (this can also be applied if you have the cDNA yourself in any vector). Please ask for more details regarding this customized service!
How can I make a clone request?
- Please fill in the clone request form and send it to us as an email attachment. Remember to add your research group, clone name and position and possible cloning service needed.
What information is available for my clone?
- The ORFeome library is validated by the vendor and we have performed random validations by sequencing. MGC library is validated by the vendor by sequencing the N-terminus of the clones.
- For checking clone sequences, search database with the accession number stated for each clone in the excel (in most cases the accession number has prefix "BC").
Should I pay attention to the stop codon status of ORFeome entry clones?
- The ORFeome collection contains both entry clones with stop codon and entry clones without stop codon. The stop codon status is stated for each clone in the excel sheet.
- If you'd like to request a cloning into an expression vector where your protein would be expressed with a C-terminal tag/epitope fusion, you need to choose an entry construct without stop codon for the cloning.
Do I need stable E. Coli cells for growing lenti constructs?
- It is recommended to grow lenti constructs in E. Coli (Stbl3) as the lentiviral DNA contains direct repeats and is less stable.
- If you need to inoculate new cultures for preparing more lentiviral DNA, transform the constructs to Stbl3 cells or use the glycerol stock obtained from GBU. Please ask for the glycerol stock at the same time you receive the constructs.
How do I get my clones?
- We provide the clones as a streak on a bacterial plate and cloned ORFs as a miniprep. We send plates/tubes by mail or you can pick them up. If requested, you can also obtain the glycerol stock when picking up the plate/miniprep, but we don't send glycerol stocks by mail.
Should I sequence my clones?
- We highly recommend sequencing the clones before using them in experiments.
- Suitable sequencing primers are stated in an info sheet you receive as email attachment when your order is ready.
- Viikki sequencing service and Biomedicum sequencing unit have primers for sequencing constructs from GBU.
What if there is an error with a clone I receive?
- The clone collections are sequence verified by the vendor. GBU has validated the libraries by random sequencing and observed some errors on the plates. On this account we highly recommend sequencing the clones before using them in experiments.
- If sequencing shows the clone you have received is not the one you requested, please contact GBU immediately. Please make sure you compare the sequencing result to the exact same sequence that is indicated in the excel sheet for your clone.
- Upon your notification on the wrong clone we will check the plate it originated from to rule out errors with picking the clone.
- If possible, incorrect clones are reimbursed by picking the same or equivalent clone from another location in the library.
- If the same or equivalent clone cannot be found in other locations in the collection, the incorrect clone will not be charged but cannot be compensated otherwise.
Can I request empty vectors from GBU e.g. for transfection controls?
- GBU does not provide empty Gateway destination vectors.
- If you need a control construct we have enhanced GFP and mCherry sequences cloned into destination vectors. Please indicate the need for a control construct in the clone request form.
Does the cloning service include the cloning of customers cDNAs and/or to customers vectors?
- Please send a request via email if you are interested in using your own constructs in cloning.
Please note: GBU clones are under MTA terms from the suppliers. See MTA terms provided with a clone. The fee is solely from transportation and packaging.