Blue:SingleCelltotoRNAseqProgress

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Revision as of 18:15, 28 March 2013 by >B1lake
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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