What is the difference between MGC and ORFeome libraries?
- ORFeome library: All clones contain open reading frames (ORFs) in Gateway‑compatible entry vectors (pENTR221, pENTR201, pENTR223). These clones are designed for straightforward recombination into a wide range of Gateway destination vectors. .
- MGC library: The clones are full‑length cDNAs in a variety of vector backbones. Because the vectors differ, you should check the specific backbone information in the library Excel file. .
How do I know whether you have the clone I'm interested in?
- You can check the availability of specific clones by browsing the Excel lists on our cDNA, shRNA, or gRNA clone collection pages. Each file includes the full set of clones currently available in that library.
What if I can't find my clone of interest?
- If you cannot locate the clone in our Excel sheets, it is unfortunately not included in our collections. Before concluding this, please ensure you have searched using all possible gene names, synonyms, and acronyms. Also note that human and mouse constructs are listed on separate sheets.
- If the clone is available only in the MGC collection but the vector is not suitable for your experiment, we can generate a Gateway‑compatible entry clone by PCR. This option is also available if you already have the cDNA in any vector. Please contact us for more details about this customized service.
- If suitable gRNAs or shRNAs are not available, we can assist with the design and cloning of .
- You may also check the websites of commercial providers such as , , or .
How can I make a clone request?
- Please fill in the clone request form and send it to us as an email attachment. Make sure to include your research group, the clone name and ID, and any cloning services you may require.
What additional information is available for my clone apart from the Excel lists?
- The ORFeome library clones are validated by the vendor, and we have additionally performed random sequence validations. MGC library clones are validated by the vendor by sequencing the N‑terminus of each construct. To check the sequence of a specific clone, search public databases using the accession number provided in the Excel sheet (see the column labeled GB_ACCNUM or Accession List).
- For shRNA constructs, detailed sequence information and validation status can be found by searching for the target gene on the manufacturer’s .
- For gRNA constructs, additional information is available by searching for the gene of interest on the Sanger CRISPR Clones .
Should I pay attention to the stop codon status of ORFeome entry clones?
- Yes. The ORFeome collection includes both entry clones with a stop codon and clones without a stop codon. The stop codon status for each construct is indicated in the Excel sheet.
- If you plan to request cloning into an expression vector that adds a C‑terminal tag or epitope, you must choose an entry clone without a stop codon. This ensures that the tag is fused in‑frame with your protein.
Do I need stable E. coli cells for growing lenti constructs?
- Yes. It is recommended to grow lentiviral constructs in stable E. coli strains (e.g., Stbl3) because lentiviral plasmids contain direct repeats and are therefore more prone to recombination in standard cloning strains.
I need more plasmid DNA of the construct I ordered from GBU — what should I do?
- If you need additional plasmid DNA, you can inoculate a new culture using the glycerol stock provided by the Genome Biology Unit or by transforming the plasmid into an appropriate E. coli strain (e.g., Stbl3 for lentiviral constructs). If you did not request a glycerol stock with your original order, please contact us so we can provide one.
How do I get my clones?
- We provide clones as a streak on a bacterial plate, as minipreps, or as glycerol stocks.
- Plates are sent only via internal mail, or you may pick them up in person.
- Cloned constructs are delivered as minipreps, and a glycerol stock is always prepared.
- DNA miniprep and glycerol stock tubes can be sent by mail, or you can collect them on site. For glycerol stocks, UPS shipment on dry ice is available (shipping fees will be added to the invoice).
Should I sequence my clones?
- The clone collections are sequence‑verified by the vendor. The Genome Biology Unit has additionally performed random sequencing and has observed occasional errors on the plates. For this reason, we highly recommend sequencing your clones before using them in experiments.
- Suitable sequencing primers are listed in the information sheet you will receive as an email attachment when your order is ready.
- If needed, you can also request clone verification from us. We offer Sanger sequencing of the insert as well as whole‑plasmid sequencing.
What if there is an error with a clone I receive?
- If sequencing shows that the clone you received is not the one indicated in the Excel sheet, please contact GBU immediately. Make sure to compare your sequencing result to the exact reference sequence listed for that clone in the Excel file.
- Once notified, we will check the original plate to rule out picking errors.
- If possible, we will replace the incorrect clone by picking an equivalent clone from another location in the library. If no equivalent clone exists elsewhere in the collection, the incorrect clone will not be charged, but we unfortunately cannot provide a replacement.
Can I request empty vectors from GBU, for example for transfection controls?
- GBU does not provide empty vectors.
- If you need a control construct, we offer several inserts (e.g., EmGFP, mCherry) cloned into destination vectors. Please indicate the need for a control construct in the order form.
Does the cloning service include cloning of customers’ cDNAs and/or into customers’ vectors?
- If you are interested in using your own constructs for cloning, please send us a request by email so we can assess feasibility.
Please note that GBU clones are distributed under the MTA terms of the suppliers of the clone collections and destination vectors. The MTA terms are provided with each clone. The fee associated with distributing clones from our collections covers transportation and packaging only.