Blue:SingleCelltotoRNAseqProgress
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Experiment | Sample | Protocol | Lysis Buffer | Fragmentation | Capping | RT Purification | TSO Vol | PCR Method | PCR Outcome | DSN treatment | Post-DSN Outcome | RNA-Seq Outcome |
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 | |||||||||
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 | |||||||||
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 | |||||||||
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 | |||||||||
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 | ||||||||
RNAseIII @ 37C 20min 65C 10min | ||||||||||||
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 | |||||||||
37C 5min | Size Selection required | |||||||||||
RNAseIII @ 37C 10min 65C 5min | ||||||||||||
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 | ||||||||||||
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 | ||||||||||||
RNAseIII @ 37C 10min 65C 10min |