Complete FISH probe sets can be efficiently labeled in bulk with comparable efficiency to commercial probes. (A) PAGE analysis of Quasar570 labeling of multiple probe sets. Probe sets A to O (lanes 3–17) contained from 15 to 48 oligonucleotides each, targeting a particular cellular transcript. Lane 2 contains the unlabeled oligos of set A. Lane 17 (set O) contains a commercial probe set from Stellaris. Band Doublets in some lanes indicate mobility differences among the mixed probes. (B) Box plots show the DOL of multiple probe sets determined by spectrophotometer and by PAGE analysis. Each dot represents one probe set. (C) The correlation of DOL measurements for Quasar570 labeling by spectrophotometer and PAGE analysis. The fitted linear equations and R2 values are indicated. The probe set from Stellaris is highlighted in red. (D) PAGE analysis of Quasar670 labeling efficiency for multiple oligo sets. (Lanes 3–13) Sets I to XI are dye-labeled oligo sets containing 15–48 oligos each. Lane 2 contains the original unlabeled oligos of set I. Lane 7 (set V) contains a commercial probe set from Stellaris. (E) Box plots of the Quasar670 DOL for the multiple probe sets determined by spectrophotometer (see Supplemental Fig. S2) and PAGE. (F) The correlation of Quasar670 DOL measurements by spectroscopy and PAGE is displayed by scatter plot. The fitted linear equations and R2 values are indicated. The probe set from Stellaris is highlighted in red. Note that these gels were run for less time than that in Figure 1, such that the labeled and unlabeled oligos are not separated as completely.
