Hosuk:LabNotes/2014-1-27: Difference between revisions
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*Not many, just a few, 10~20 from 2nd rolonies. | *Not many, just a few, 10~20 from 2nd rolonies. | ||
*Need positive control or some information about expected abundance of each gene. | *Need positive control or some information about expected abundance of each gene. | ||
[[File:NCIH1975_Mutant_dcProbeCount.png| | [[File:NCIH1975_Mutant_dcProbeCount.png|500px]] | ||
Revision as of 02:06, 28 January 2014
NCIH1975 : Mutation Detection
- Continued from 1/19
2nd Rolony
- Annealing Padlock probes, Ampligase, and 2nd RCA fab. --> done at 01/19~01/21
Detection of Mutation
- Procedure
- Add Cy5-Adapt to detect 1st Rolonies, 3 positions
- Strip (80% Formamide for 15min), Add dcProbe1 (incubate 15min), image
- Strip (80% Formamide for 15min), Add dcProbe2 (incubate 15min), image
- Strip (80% Formamide for 15min), Add dcProbe3 (incubate 15min), image
- Strip (80% Formamide for 15min), Add dcProbe4 (incubate 15min), image
Result
- Image at 3 different positions
1st Rolony
- Pos1 : 5151
- Pos2 : 5964
- Pos3 : 6485
File:NCIH1975 Mutant Pos1-2-3 1stRolony.png
Each dcPobes
- Not many, just a few, 10~20 from 2nd rolonies.
- Need positive control or some information about expected abundance of each gene.
File:NCIH1975 Mutant dcProbeCount.png
- Images of each gene at each position
- Circles are the 2nd Rolony that I've counted.
File:NCIH1975 Mutant Pos1 dcProbe.png
File:NCIH1975 Mutant Pos2 dcProbe.png