PCR was carried out for the screening of the cells harboring the recombinant the pHT01-kIspS plasmid using the same primers and amplification conditions as mentioned above.
In the 3rd (final) screening of the cells in the flask, the cells showed high reactivity to V3 (OD > 1) (Table 2).
Beginning with the 96 well stage and up to the final screening of the cells in the T25 flask, we selected only those wells with high V3 binding reactivity for further expansion and finally for the phage library construction.
Therefore, we consider this analysis a preliminary screen of the cell cycle pathway and one which indicates modest evidence for association with disease risk for only one gene, ABL1.
Primer 9 was used in place of primer 4 in the initial screening of the target cell lines.
Whole mitochondrial genome screening of the six cell lines generated 114 amplicons representing around 100 kb of DNA.
We next sought to confirm that the sensitivity/resistant phenotype observed during screening of the HOMUR cell lines was reproducible.
We screened all of the cell populations by FACS except for the differentiated NSC, which we screened by image to take advantage of the distinct morphological differences between neurons and non-neuronal cells.
In search of other possible S phase checkpoint defects, we screened all of the cell lines in the NCI60 panel for their ability to S-phase arrest in response to the topoisomerase I inhibitor SN38.
Screening of the SKP cells from dissociated HGPS spheres indicated that an average of 14%to26%6% of the cells exhibited a detectable progerin-positive signal, whereas the normal SKPs showed no progerin signal (Fig. 5A).
