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== Smart-Seq2 Library Preparation == :'''TSO Reaction''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |Purified cDNA||||||||||3ul |- |5x First Strand buffer||||||||||2ul |- |10mM dNTP mix||||||||||1ul |- |100mM DTT||||||||||0.5ul |- |100uM Smart2-TSO||||||||||0.1ul |- |MgCl2 (1M)||||||||||0.06ul |- |Betaine (5M)||||||||||2ul |- |RNase Inhibitor||||||||||0.25ul |- |SuperScriptII||||||||||0.5ul |- |dH2O||||||||||0.59ul |- |Total||||||||||10ul |- |} :::''Incubate @42C 20min'' :::''Incubate [@50C 2min > 42C 2min] x10 cycles'' :::''Incubate @70C 10min'' :::''Incubate @10C hold'' :'''PCR (KAPA-HiFi)''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |TSO Reaction||||||||||10ul |- |2x KAPA HiFi Hotstart ReadyMix||||||||||12.5ul |- |IS PCR Primer (10uM)||||||||||0.25ul |- |H2O||||||||||2.25ul |- |Total||||||||||25ul |- |} ::{| {{table}} |- |98C 3min|||||||||| |- |98C 20s -> 67C 15s -> 72C 1min||||||||||15x (1ng); 18x (100pg); 21x (10pg) |- |72C, 5min |- |}
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