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The fold increase in exogenous ezrin expression was determined by comparing the titration end-point of the corresponding ezrin band in each clone with that of cells transfected with empty pCB6 vector.
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Western blot analysis demonstrated a 170 kDa band with each clone.
Six clones per unique band in a clone fingerprint was expected, yet regions of 12 and 24 clones per unique band were common.
The average number of fragments (bands) of each clone was 69.4.
The average number of DNA bands generated from each clone was 41 bands on a calculation using the whole FPC database.
We estimated the sizing accuracy of the capillary sequencer by analyzing the size of the vector band in 200 clones.
The average number of DNA bands generated from each clone was 40 on a calculation from the 4,754 FPC database.
The product of TNFRSF11A_Δ7,8,9 variant in this study was named RANK-c, because it lacks three exons and was identified as the third and smaller migrating band in our initial cloning experiments.
6 DNA fragments with length from 315 to 4557 bp were successfully cloned in the validation experiments, and the 100% positive clone rate of 10 randomly-picked clones in each cloning experiment was verified by means of colony PCR.
Clone A12 DNA also gave single major bands in each of the two digests with sizes distinct from those in both Clone A1 and the plasmid control, indicating the presence of a different single integration site in this clone.
Tolerance determines how closely two bands in different clones need to match to be considered as the same band.
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