Hosuk:LabNotes/2014-4-10
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Cancer Cell : Mutation detection test - 2nd Try
- NCIH-1975 in 24 well plate
- Fix : 04/08
- Finish all process 04/10
- Well 1, 2 : EGFR, BRAF RT Primers
- Well 3, 4 : EGFR, BRAF RT Primers + BRAF Stop primer 200bp
- Well 5, 6 : EGFR, BRAF RT Primers + BRAF Stop primer 200bp
- Well 7, 8, 9, 10 : random hexamer
File:24Well 2014-04-10.png
File:NCIH1975 2014-04-10 1stRolony Count.png
Result for 1st Rolonies
- Well 1 ~ Well 6 have similar number of rolonies,
- which the numbers are much less than Well 7, 8, and 9.
- (Well 10 has few, it’s because of mistake at RNaseH step)
PGP1F sample for 13k Padlock probe test
- Fix : 04/06, 2 dishes
- Finish all process 04/10, imaging
- S1 : incubate RCA primer at RT for 10min after incubating RCA primer at 60C oven for 15min.
- S2 : Wash and add RCA mix right after incubating RCA primer at 60C oven for 15min.
Result for 1st Rolonies
- Both S1 (incubate RCA primer at RT for 10min), S2 looked similar amount of Rolonies
- Both sample have pretty good 1st Rolonies!
- I will use S2 for Ampligase and RCA with13k Padlock probes.