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 |
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*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: | ||
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| 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 | |||
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|- | |||
| 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 | |||
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== 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 |