Blue:RNA-Seq Experiments:07282014: Difference between revisions

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>B1lake
>B1lake
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* Yield for the RNA is too high (expect more of that seen for the 100-500 nuclei), therefore repeated PCR using 3 less cycles (18 total)

Revision as of 00:28, 30 July 2014

SmartPlus on Mouse Nuclei

Obtained sorted nuclei from Aran:

  1. 100 nuclei sorted directly into 2ul clontech dilution buffer (4x)
  2. 500 nuclei sorted directly into 2ul clontech dilution buffer (4x)
  3. 5000 nuclei sorted into BSA buffer (2x)
  • stored 100/500 sorts at -80C. Vacuum spun down to 1ul for each.
  • RNA extraction of 5000 using Zymo Micro RNA Kit (including DNAse treatment). Eluted in 6ul and vacuum spun down to 2ul.
  • Performed SmartPlus protocol as per C1 tube control protocol on 1ul of above samples (only modification is addition of randomer) using 21 cycles for PCR
  • cDNA Yields:
Sample Sample Concentration (ng/ul)
NC 0.999
RNA 1 72.1
RNA 2 40.0
100 nuc 1 7.93
100 nuc 2 8.75
500 nuc 1 5.20
500 nuc 2 6.25


  • Yield for the RNA is too high (expect more of that seen for the 100-500 nuclei), therefore repeated PCR using 3 less cycles (18 total)