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*[[Hosuk:Lab_Notes|LabNote]] ===Rolony, 9th=== *Try the method that I’ve learned from Jonathan and Jay *2uL Riboshredder + 10uL Rnase H + 20uL Rnase H buffer + 168uL H2O (Jonathan’s protocol) *Use double volume of CircLigase reagent except CircLigase II (Jonathan’s protocol) *Use Enzymatics’ Phi29 --> 1uL Phi29 (High concentration of Enzymatics) for RCA mix *Use plastic bag with wet tissue for long time incubation *Start at 06/16 *Use cell dishes fixed at 06/04 *S3 --> -RT, S4 --> +RT *Run RT : 9:00pm 06/16 ~ 9:00am 06/17 --> 12hr. *Run CircLigase II : 12:05am 06/17 ~ 4:05pm 06/17 --> 4hr. *Run RCA : 5:00 pm 06/17 ~ 3: 10pm 06/18 --> 22hr. ====Result==== *S3, -RT sample didn't show any rolony signal. *S4, +RT sample: had not as many rolonies as what I expected, but not bad! **Low background, and no signal through FITC nor Cy5 channel though, ====epi FL==== *'''-RT, 20x, NA=0.8''' [[File:S3_-RT_20x-NA0.8_pos02.png|600px]] *'''+RT, 20x, NA=0.8''' [[File:S4_+RT_20x-NA0.8_pos02.png|600px]] *'''+RT, 40x Oil, NA=1.3''' [[File:S4_+RT_40xOil-NA1.3_pos03.png|600px]]
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