Blue:RNA-Seq Experiments:09232013: Difference between revisions

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>B1lake
(Created page with "== 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 *Centrifuge...")
 
>B1lake
Line 8: Line 8:
*Add 15ul of nuclei to 10ul cell suspension reagent and loaded 20ul onto chip
*Add 15ul of nuclei to 10ul cell suspension reagent and loaded 20ul onto chip
*Observed chip under microscope and counted occupancy rate:
*Observed chip under microscope and counted occupancy rate:
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1'''
| align="center" style="background:#f0f0f0;"|'''2'''
| align="center" style="background:#f0f0f0;"|'''3'''
| align="center" style="background:#f0f0f0;"|'''4'''
| align="center" style="background:#f0f0f0;"|'''5'''
| align="center" style="background:#f0f0f0;"|'''6'''
| align="center" style="background:#f0f0f0;"|'''7'''
| align="center" style="background:#f0f0f0;"|'''8'''
| align="center" style="background:#f0f0f0;"|'''9'''
| align="center" style="background:#f0f0f0;"|'''10'''
| align="center" style="background:#f0f0f0;"|'''11'''
| align="center" style="background:#f0f0f0;"|'''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 ==

Revision as of 01:27, 24 September 2013

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