Hosuk:LabNotes/2014-4-10: Difference between revisions
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[[File:24Well_2014-04-10.png| | [[File:24Well_2014-04-10.png|300px]] | ||
[[File:NCIH1975_2014-04-10_1stRolony_Count.png|150px]] | |||
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*which the numbers are much less than Well 7, 8, and 9. | *which the numbers are much less than Well 7, 8, and 9. | ||
**(Well 10 has few, it’s because of mistake at RNaseH step) | **(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. | |||
[[File:PGP1F_2014-04-10_1stRolony_Count.png|400px]] |
Latest revision as of 21:51, 10 April 2014
Cancer Cell : Mutation detection test - 2nd Try[edit]
- 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[edit]
- 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[edit]
- 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[edit]
- 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.