Matt:LabNotes/2014-1-31: Difference between revisions

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>Mzcai
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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