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===Mutation Detection with Mixed with PGP1F and NCIH1975 : Ampligase, 2nd RCA=== *Continied from [[Hosuk:LabNotes/2014-3-6|03/06]] ====Experiment==== *8 Ampligase reaction, *4 NTC(no Ampligase emzyme) [[File:Mix_PGP1F_NCIH1975_24wellPlates_AmpligaseMap.png|300px]] *Procedure *#Make Ampligase mix, add *#Incubate at 45C for 24hr. (03/13 ~ 03/14) *#Run RCA reaction for 24hr. (03/14 ~ 03/15) *#Anneal dcProbes, and Image *Ampligase Mix **10x Buffer 20uL **Ampligase (5U/uL) 8uL **ppEGRF_T790M_WT 2uL **ppEGRF_T790M_MU 2uL **ppEGRF_L858R_WT 2uL **ppEGRF_L858R_MU 2uL **H2O 164 uL *NTC Mix **10x Buffer 20uL **Ampligase (5U/uL) 0uL **ppEGRF_T790M_WT 2uL **ppEGRF_T790M_MU 2uL **ppEGRF_L858R_WT 2uL **ppEGRF_L858R_MU 2uL **H2O 172 uL ====Result : 1st Rolonies==== *Number of rolonies from most of wells are not bad. *PGP1F has more 1st rolonies than cancer cell has. **I think main reason is the size of PGP1F is much more than cancer cell's, which make a big different. **That means the density per '''area''' of rolonies seems similar at each cell type. [[File:Mix_PGP1F_NCIH1975_Counts.png|500px]] [[File:Mix_PGP1F_NCIH1975_Well04.png|800px]] [[File:Mix_PGP1F_NCIH1975_Well06.png|800px]] [[File:Mix_PGP1F_NCIH1975_Well10.png|800px]]
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