I have obtained EST clones from the IMAGE consortium (from RLDB in
Berlin). According to the database information these clones (0.4 and 0.5 kb inserts) are in pCMV-Sport2 in DH10B.
I have streaked out the bacteria on 2TY/50 microg/ml AMPICILLIN
plates and picked 15 colonies of each clone. RE-mapping with
NotI/SalI and XhoI/EcoRI showed a 4 kb band but NO insert.
PCR with T7 and SP6 primers also failed to amplify a specific product
from the crude bacterial stab (same with specific primers for one of
the clones). Did I overlook something? Has anybody
Have a nice weekend
Dr. Fabian F. MOEBIUS
Institut fuer Biochemische Pharmakologie
Peter Mayr Str. 1