Blue:RNA-Seq Experiments:09232013

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Fluidigm C1 Run: Human Pyramidal Nuclei

  • Samples prepared and sorted by Yun at Scripps (FACS ARIA)
  • Bulk sorted nuclei based on PI positive - ~120 nuclei/ul
  • Centrifuged nuclei at 2000rpm 4C for 10 min (table-top centrifuge)
  • Resuspended in low volume and counted - ~250 nuclei/ul
  • Used ERCC diluted 1:200,000 (10 fold less than previous experiment)
  • Add 15ul of nuclei to 10ul cell suspension reagent and loaded 20ul onto chip
  • Observed chip under microscope and counted occupancy rate:
' 1 2 3 4 5 6 7 8 9 10 11 12
A 3_C03 1_C02 0_C01 0_C49 0_C50 0_C51 0_C06 0_C05 0_C04 0_C52 0_C53 1_C54
B 0_C09 1_C08 2_C07 0_C55 1_C56 0_C57 0_C12 0_C11 0_C10 0_C58 0_C59 1_C60
C 0_C15 1 frag_C14 0_C13 1_C61 1_C62 0_C63 0_C18 1_C17 1_C16 0_C64 0_C65 1_C66
D 1_C21 0_C20 1 + 1 frag_C19 0_C67 2_C68 0_C69 0_C24 1_C23 6_C22 0_C70 0_C71 0_C72
E 1_C25 0_C26 0_C27 0_C75 0_C74 0_C73 3_C28 1_C29 0_C30 0_C78 0_C77 1_C76
F 0_C31 0_C32 0_C33 2_C81 0_C80 0_C79 1 + 1 frag_C34 0_C35 0_C36 0_C84 0_C83 1_C82
G 3_C37 0_C38 1_C39 0_C87 0_C86 0_C85 0_C40 1_C41 0_C42 0_C90 1_C89 0_C88
H 0_C43 4_C44 0_C45 1_C93 0_C92 0_C91 1_C46 3_C47 6_C48 0_C96 0_C95 0_C94


RNA Extraction of Unfixed and Fixed Nuclei