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*[[Hosuk:Lab_Notes|LabNote]] ===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 β 4 probes *#*EGFR T790M(Wild, Mutant), EGFR L858R(Wild, Mutant) *#*50nM each in 2x SSC + 30% Formamide (2uL each) *#*Preheat probes at 80C for 5min, incubate at 50C for 4hr. *#Add SNL mix (Ampligase + KlenTaq enzyme mix) *#Incubate 50C for 20hr. *#Anneal Primers for 2nd RCA *#Run RCA reaction for overnight(20hr.) *#Anneal dcProbes, and Image [[File:ReactionMix_Ampligase_KlenTaq.png|600px]]
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