– 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 is less necessary when you subclone the ORF into another vector, as it implies a colony-picking step and sequencing step anyway.
– 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.