Blue:RNA-Seq Experiments:09052013: Difference between revisions
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*Achieved good loading efficiency (81/96) but high mortality as indicated by live/dead staining on chip | *Achieved good loading efficiency (81/96) but high mortality as indicated by live/dead staining on chip | ||
*Followed the protocol exactly - including use of spike-in RNA indicated in the protocol rather than those used in our previous experiments | *Followed the protocol exactly - including use of spike-in RNA indicated in the protocol rather than those used in our previous experiments | ||
'''OutCome:''' | |||
*Re-primed incorrectly - broke seal on chip preventing lysis buffer from entering chambers - no cDNA amplification | |||
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[[File:Data_Submission_Worksheet.docx]] | [[File:Data_Submission_Worksheet.docx]] | ||
== Sample Attribution == | |||
Note: d=dead | |||
{| {{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||1_NPCs_C03||0_NPCs_C02||1d_NPCs_C01||0_NPCs_C49||1d_NPCs_C50||1d_NPCs_C51||1d_NPCs_C06||0_NPCs_C05||1_NPCs_C04||2d_NPCs_C52||0_NPCs_C53||3d_NPCs_C54 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| B||1d_NPCs_C09||0_NPCs_C08||0_NPCs_C07||1d_NPCs_C55||1d_NPCs_C56||1d_NPCs_C57||1d_NPCs_C12||1d_NPCs_C11||1d_NPCs_C10||1d_NPCs_C58||0_NPCs_C59||4d_NPCs_C60 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| C||1d_NPCs_C15||1d_NPCs_C14||0_NPCs_C13||1_NPCs_C61||0_NPCs_C62||1d_NPCs_C63||1_NPCs_C18||1_NPCs_C17||1_NPCs_C16||1d_NPCs_C64||1d_NPCs_C65||1d_NPCs_C66 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| D||1d_NPCs_C21||1d_NPCs_C20||0_NPCs_C19||1_NPCs_C67||1d_NPCs_C68||1d_NPCs_C69||2d_NPCs_C24||1d_NPCs_C23||1d_NPCs_C22||0_NPCs_C70||1d_NPCs_C71||1 + 1d_NPCs_C72 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| E||1d_NPCs_C25||3d_NPCs_C26||1_NPCs_C27||0_NPCs_C75||1d_NPCs_C74||0_NPCs_C73||2d_NPCs_C28||2d_NPCs_C29||1d_NPCs_C30||0_NPCs_C78||1d_NPCs_C77||2 + 1d_NPCs_C76 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| F||4d_NPCs_C31||1_NPCs_C32||1d_NPCs_C33||1_NPCs_C81||1d_NPCs_C80||2d_NPCs_C79||1d_NPCs_C34||1d_NPCs_C35||3 + 1d_NPCs_C36||1d_NPCs_C84||1_NPCs_C83||1_NPCs_C82 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| G||1d_NPCs_C37||0_NPCs_C38||1_NPCs_C39||1d_NPCs_C87||2d_NPCs_C86||1 + 1d_NPCs_C85||1_NPCs_C40||1_NPCs_C41||1d_NPCs_C42||2d_NPCs_C90||1_NPCs_C89||1_NPCs_C88 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| H||1d_NPCs_C43||4d_NPCs_C44||1d_NPCs_C45||1d_NPCs_C93||1d_NPCs_C92||4d_NPCs_C91||3d_NPCs_C46||1d_NPCs_C47||1d_NPCs_C48||1d_NPCs_C96||1d_NPCs_C95||1d_NPCs_C94 | |||
|- | |||
| |||||||||||||||||||||||| | |||
|- | |||
| | |||
|} |
Latest revision as of 16:31, 11 October 2013
Fluidigm C1 Small Chip Beta Test[edit]
- Tested NPCs on the small 5-10um chip
- First run on 09-04-2013 produced too few viable cells for loading and resulted in only 10 captured - therefore, reprimed the chip and stored overnight for re-loading
- Second run on 09-05-2013 involved E13.5 neural progenitors sorted as PI negative
- Achieved good loading efficiency (81/96) but high mortality as indicated by live/dead staining on chip
- Followed the protocol exactly - including use of spike-in RNA indicated in the protocol rather than those used in our previous experiments
OutCome:
- Re-primed incorrectly - broke seal on chip preventing lysis buffer from entering chambers - no cDNA amplification
File:Cell Characterization Worksheet.docx
File:Data Submission Worksheet.docx
Sample Attribution[edit]
Note: d=dead
' | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
A | 1_NPCs_C03 | 0_NPCs_C02 | 1d_NPCs_C01 | 0_NPCs_C49 | 1d_NPCs_C50 | 1d_NPCs_C51 | 1d_NPCs_C06 | 0_NPCs_C05 | 1_NPCs_C04 | 2d_NPCs_C52 | 0_NPCs_C53 | 3d_NPCs_C54 |
B | 1d_NPCs_C09 | 0_NPCs_C08 | 0_NPCs_C07 | 1d_NPCs_C55 | 1d_NPCs_C56 | 1d_NPCs_C57 | 1d_NPCs_C12 | 1d_NPCs_C11 | 1d_NPCs_C10 | 1d_NPCs_C58 | 0_NPCs_C59 | 4d_NPCs_C60 |
C | 1d_NPCs_C15 | 1d_NPCs_C14 | 0_NPCs_C13 | 1_NPCs_C61 | 0_NPCs_C62 | 1d_NPCs_C63 | 1_NPCs_C18 | 1_NPCs_C17 | 1_NPCs_C16 | 1d_NPCs_C64 | 1d_NPCs_C65 | 1d_NPCs_C66 |
D | 1d_NPCs_C21 | 1d_NPCs_C20 | 0_NPCs_C19 | 1_NPCs_C67 | 1d_NPCs_C68 | 1d_NPCs_C69 | 2d_NPCs_C24 | 1d_NPCs_C23 | 1d_NPCs_C22 | 0_NPCs_C70 | 1d_NPCs_C71 | 1 + 1d_NPCs_C72 |
E | 1d_NPCs_C25 | 3d_NPCs_C26 | 1_NPCs_C27 | 0_NPCs_C75 | 1d_NPCs_C74 | 0_NPCs_C73 | 2d_NPCs_C28 | 2d_NPCs_C29 | 1d_NPCs_C30 | 0_NPCs_C78 | 1d_NPCs_C77 | 2 + 1d_NPCs_C76 |
F | 4d_NPCs_C31 | 1_NPCs_C32 | 1d_NPCs_C33 | 1_NPCs_C81 | 1d_NPCs_C80 | 2d_NPCs_C79 | 1d_NPCs_C34 | 1d_NPCs_C35 | 3 + 1d_NPCs_C36 | 1d_NPCs_C84 | 1_NPCs_C83 | 1_NPCs_C82 |
G | 1d_NPCs_C37 | 0_NPCs_C38 | 1_NPCs_C39 | 1d_NPCs_C87 | 2d_NPCs_C86 | 1 + 1d_NPCs_C85 | 1_NPCs_C40 | 1_NPCs_C41 | 1d_NPCs_C42 | 2d_NPCs_C90 | 1_NPCs_C89 | 1_NPCs_C88 |
H | 1d_NPCs_C43 | 4d_NPCs_C44 | 1d_NPCs_C45 | 1d_NPCs_C93 | 1d_NPCs_C92 | 4d_NPCs_C91 | 3d_NPCs_C46 | 1d_NPCs_C47 | 1d_NPCs_C48 | 1d_NPCs_C96 | 1d_NPCs_C95 | 1d_NPCs_C94 |