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Clones selected for both the BAC and the shuttle vector were then grown at 43 °C, the temperature at which the shuttle vector's origin of replication is nonfunctional, thus forcing continued antibiotic selection to occur on a single molecule.
We therefore refer to genes that meet this cutoff as "gene dependencies". Screens performed in cell clones selected for optimal editing identified most gene dependencies (Fig. 1c and below).
All the sequences obtained from 5 clones selected for each protozoan matched 100% with the GenBank sequences.
These were then used to transform electrocompetent S. aureus RN4220, according to the method of Schenk and Ladagga [80], with clones selected for on the basis of tetracycline and erythromycin resistance.
In fact, only one of the 19 TAR clones selected for sequencing (TAR128) is inconsistent with other data and could be a recombinant between two BESs generated during TAR cloning.
(XLSX 40 kb) Additional file 6: The addresses of the BAC clones selected for sequencing.
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The bovine SA clone selected for sequencing was isolated from a cow presenting with clinical mastitis in 1993 [12].
The rosetting clone 3D7S8.4 transcribed the var-gene PFD0630c at onset whilst the 3D7AH1S2 clone selected for CD36 adhesion transcribed PFF0845c.
BT474-HR6 clone, selected for its resistance to the ErbB-2 antibody trastuzumab, was already described [ 27] and was a gift from C. Arteaga (Vanderbilt University, Nashville, TN, USA).
After dot-blot and elimination of the most likely repetitive clones, 753 clones were selected for sequencing.
Amplicons were cloned and totally 479 clones were selected for sequencing.
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