Hosuk:LabNotes/2014-1-19: Difference between revisions
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>Hosuki78 (Created page with "*LabNote ===NCIH1975 : Mutation Detection=== ====Padlock Probes==== *Padlock probes selection *1)EGFR T790M **Wild : detected by dcProbe1 **Mutant : detec...") |
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*#Anneal dcProbes, and Image | *#Anneal dcProbes, and Image | ||
[[File:ReactionMix_Ampligase_KlenTaq.png|550px]] |
Revision as of 00:35, 20 January 2014
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 70C for 5min, incubate at 50C for 1hr.)
- Add SNL mix (Ampligase + Stoffel enzyme mix)
- Incubate 50C for 4hr.
- Anneal Primers for 2nd RCA
- Run RCA reaction for overnight(12hr.)
- Anneal dcProbes, and Image