Matt:LabNotes/2014-1-31: Difference between revisions
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| ||||||Average % of Cy3:||51.04 | | ||||||Average % of Cy3:||51.04 | ||
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====Conclusions==== | |||
*If we take the very rough estimate that 51% of primary rolonies are hybridized by Shifted FISSEQ dye | |||
**Previously when we hybridized ppMALAT1 and found 59% of primary rolonies are MALAT1, then the '''true''' number of MALAT1 rolonies is 59-51 = ~8% | |||
===RT_RAB7A Primers and the Number of RAB7A Primary Rolonies=== | ===RT_RAB7A Primers and the Number of RAB7A Primary Rolonies=== |
Revision as of 22:17, 6 February 2014
Shifted FISSEQ vs FISSEQ_Adpt Signal Count in Same Wells
- Previously compared the two dyes in adjacent wells Matt:LabNotes/2014-1-28
- Here I strip FISSEQ_Adpt from before and add Shifted FISSEQ to column 9
- Put Shifted FISSEQ_2ndRCAprimerDye in columns 4 and 9
- Put RAB7A_1stRolonyFISH in columns 3 and 10
Well | RT primer | Cy3 | Shifted | % of Cy3 |
C9 | 220nt | 711 | 129 | 18.14 |
D9 | 300nt | 357 | 203 | 56.86 |
E9 | 510nt | 4449 | 842 | 18.923 |
F9 | 1000nt | 13295 | 14654 | 110.22 |
Average % of Cy3: | 51.04 |
Conclusions
- If we take the very rough estimate that 51% of primary rolonies are hybridized by Shifted FISSEQ dye
- Previously when we hybridized ppMALAT1 and found 59% of primary rolonies are MALAT1, then the true number of MALAT1 rolonies is 59-51 = ~8%
RT_RAB7A Primers and the Number of RAB7A Primary Rolonies
- Using RAB7A_1stRolonyFISH (ATTO550)
C10_550: 24 D10_550: 24 E10_550: 13 F10_550: 35