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== Smart-seq 2 Protocol == :'''RNA''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |UHRR/ERCC ||||||||||2ul |- |10uM T30||||||||||1ul |- |dNTP (10mM)||||||||||1ul |- |Total||||||||||4ul |- |} :::''Incubate @ 72C 3 min'' :::''Transfer to ice'' :'''Reverse Transcription''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |5xRT Buffer||||||||||2ul |- |100mM DTT||||||||||0.25ul |- |Betaine (5M)||||||||||2ul |- |10uM TSO||||||||||1ul |- |RNase Inhibitor (40U/ul)||||||||||0.25ul |- |Smarter MMLV (200U/ul)||||||||||0.5ul |- |Total||||||||||10ul |- |} :::''Incubate @ 42C 90 min'' :::''Incubate @ 70C 10 min'' :'''PCR (Advantage)''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |10x Advantage 2 PCR Buffer||||||||||5ul |- |dNTP Mix (10mM)||||||||||2ul |- |STRT Primer (10uM)||||||||||2ul |- |50X Advantage 2 Polymerase Mix||||||||||2ul |- |H2O||||||||||29ul |- |Total||||||||||50ul |- |} ::{| {{table}} |- |95C 1 min|||||||||| |- |95C 15s -> 65C 30s -> 68C 6min||||||||||12x |- |72C, 10min |- |}
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