Hosuk:LabNotes/2014-1-19
Jump to navigation
Jump to search
NCIH1975 : Mutation Detection
Padlock Probes
- Padlock probes selection
- 1)EGFR T790M
- Wild : detected by dcProbe1
- Mutant : detected by dcProbe3 or dcProbe4
- 2)EGFR L858R
- Wild : detected by dcProbe4
- Mutant : detected by dcProbe2
Procedure
- Experiment Plan
- Anneal Padlock probes – 2 probes (Preheat probes at 80C for 5min, incubate at 50C for 4hr.)
- Add SNL mix (Ampligase + Stoffel enzyme mix)
- Incubate 50C for 20hr.
- Anneal Primers for 2nd RCA
- Run RCA reaction for overnight(20hr.)
- Anneal dcProbes, and Image