Blue:SingleCelltotoRNAseqProgress: Difference between revisions

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| align="center" style="background:#f0f0f0;"|'''Experiment'''
| align="center" style="background:#f0f0f0;"|'''Sample'''
| align="center" style="background:#f0f0f0;"|'''Protocol'''
| align="center" style="background:#f0f0f0;"|'''Lysis Buffer'''
| align="center" style="background:#f0f0f0;"|'''Fragmentation'''
| align="center" style="background:#f0f0f0;"|'''Capping'''
| align="center" style="background:#f0f0f0;"|'''RT Purification'''
| align="center" style="background:#f0f0f0;"|'''TSO Vol'''
| align="center" style="background:#f0f0f0;"|'''PCR Method'''
| align="center" style="background:#f0f0f0;"|'''PCR Outcome'''
| align="center" style="background:#f0f0f0;"|'''DSN treatment'''
| align="center" style="background:#f0f0f0;"|'''Post-DSN Outcome'''
| align="center" style="background:#f0f0f0;"|'''RNA-Seq Outcome'''
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| 2272013||SSEA4+ huES9 sorted 02-26-2013||Mg||1xHM||94C 2 minutes||Yes||ETOH ppt||1x10ul||Advantage||HMW smear||1 hour Hyb||No amplification||Not Sent
|-
| ||||||(0.1%triton X-100; Rnase Inhib; ERCC)||||||||||(16 cycles)||(low yield)||||No yield||
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| 2282013||SSEA4+ huES9 sorted 02-26-2013||RNaseIII||1xRNaseIII buffer||RNAseIII @ 37C 10min 65C 10min||Yes||ETOH ppt||1x10ul||Advantage||HMW smear||1 hour Hyb||No amplification||Not Sent
|-
| ||||||(0.1%triton X-100; Rnase Inhib; ERCC)||||||||||(16 cycles)||(v. low yield)||||No yield||
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| 3032013||SSEA1+ mESCs sorted 02-07-2013||Mg||2.5xRT buffer||94C 3 minutes||No||ETOH ppt||1x10ul||Advantage||HMW smear shifted lower||1 hour Hyb||No amplification||Not Sent
|-
| ||||||(0.1%triton X-100; Rnase Inhib; ERCC)||||||||||(16 cycles)||(low yield)||||No yield||
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| 3112013||huES9 SSEA4+ sorted 02-07-2013||Mg||2.5xRT buffer||94C 5 minutes||No||Beads||1x10ul||Advantage||HMW tight smear||1 hour Hyb||No amplification||Not Sent
|-
| ||||||(0.1%triton X-100; Rnase Inhib; ERCC)||||||||||(15 cycles)||(high yield)||||No yield||
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| 3122013||SSEA4+ huES9 sorted 02-26-2013||RNaseIII||1xRNaseIII buffer||T20V.ID @72C 2min||No||Beads||2x10ul||Advantage||Correct Smear Range||1 hour Hyb||No amplification||Not Sent
|-
| ||||||(0.1%triton X-100; Rnase Inhib; ERCC)||37C 5min||||||||(15 cycles)||(v. low yield)||||No yield||
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| ||||||||RNAseIII @ 37C 20min 65C 10min||||||||||||||||
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| ||||||||||||||||||||||||
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| 3192013||Nestin mCherry High sorted 03-13-2013||RNaseIII||Hypotonic Buffer (0.2% Triton X-100; 0.5U/ul Rnase Inh)||72C 8 min||No||Beads||2x10ul||Advantage||HMW broad smear||1 hour Hyb||No amplification||Failed seq (high C content)
|-
| ||||||||T20V.ID/RNaseIII Buffer @72C 2min||||||||(15 cycles)||(high yield)||||Low yield||
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| ||||||||37C 5min||||||||||||||Size Selection required||
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| ||||||||RNAseIII @ 37C 10min 65C 5min||||||||||||||||
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| ||||||||||||||||||||||||
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| 3212013||Nestin mCherry High sorted 03-13-2013||RNaseIII||Hypotonic Buffer (0.2% Triton X-100; 0.5U/ul Rnase Inh)||72C 8 min||No||Beads||2x10ul||Advantage (15 cycles)||HMW broad smear slightly shifted lower (high yield)||1 hour Hyb||No amplification||Not sent
|-
| ||||||||T20V.ID/RNaseIII Buffer @72C 2min||||||||||||||Low yield||
|-
| ||||||||37C 5min||||||||||||||Size Selection required||
|-
| ||||||||RNAseIII @ 37C 20min 65C 5min||||||||||||||||
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| ||||||||||||||||||||||||
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| 3252013||UHRR (1ng)||RNaseIII||Hypotonic Buffer (no additives)||72C 8 min||No||Beads||1x10ul||Sybr (16 cycles)||Correct smear (high yield)||-||-||-
|-
| ||||||||T20V.ID/RNaseIII Buffer @72C 2min||||||||||||||||
|-
| ||||||||37C 5min||||||||||||||||
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| ||||||||RNAseIII @ 37C 10min 65C 10min||||||||||||||||
|}

Revision as of 18:37, 28 March 2013