– The clones are either in kanamycin-, ampicillin- or spectinomycin-resistant vectors. Some information regarding these antibiotic are provided below.
- Kanamycin: The suppliers of the ORFeome Collaboration collection recommends 30ug/ml, but your preferred concentration will also work.
- Ampicillin: It is typically used at 50-100ug/ml.
- Spectinomycin: It is typically used at 100ug/ml. We can provide some if you don’t have any. Streptomycin also works on these clones.
– Recommended growth temperatures are 30° for the lentiviral vectors clones, and 37° for other clones.
– It may be advisable to select a single colony to insure purity of the clone (despite our utmost care, tiny well-to-well contamination may always occur during handling of the clones in 96w plates format). This precaution may not be necessary in case you intend to subclone the ORF in another vector, which would most likely imply a colony-picking step and sequencing.
– We always recommend that the DNA prep of the ORFs be confirmed by sequencing to avoid any downstream problem. This is particularly true for the CCSB-Broad Lentiviral Expression collection since the final clones were not sequenced by their creators (more details here). When a clone was fully sequenced at GECF, it is indicated in the relevant database.