Point mutation detection by Hras allele specific FRET probes in NAR, 308 and MEF cells.

Abstract

<p>A - anchor<b>/</b>A,T <b>-</b> allele specific probe<b>/</b>A,U synthetic target oligo or no (O) target. After oligos transfection, cells were suspected to Acceptor Photobleach (AP) analysis. NAR cells containing Anchor and wt (A) probe showed relatively high FRET with full matched A synthetic target (blue left bar). A single mismatch in target sequence showed no FRET (U, red bar). When no target was present in the cells (NAR, green bar), no FRET was observed, indicating no FRET background. WT specific (A) probe induced a positive FRET response in both 308 and MEF cells, indicating presence of the wt allele (308– purple bar, MEF - orange bar). Mutant T probe exhibited positive FRET response only in 308 mutated cells (turquoise) as opposed to wt MEF cells (light blue).</p