Kun:LabNotes/ExonomeSeq/2007-12-21

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Revision as of 20:23, 21 December 2007 by KunZhang (talk | contribs) (New page: ==Exp. 1 Perform another round of E55kv1 capturing experiment== ===Shearing template DNA=== 10ul 353ng/ul GM10835 gDNA + 750ul shearing buffer (1X TE w/ 10% Glycerol) -> sheared with Nebul...)
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Exp. 1 Perform another round of E55kv1 capturing experiment

Shearing template DNA

10ul 353ng/ul GM10835 gDNA + 750ul shearing buffer (1X TE w/ 10% Glycerol) -> sheared with Nebulizer with unknown pressure for 1min -> recovered 650ul -> isopropanol precipitation with the presence of 4ul linear acrylamide.

Resuspended in 10ul ddH2O -> ~300ng/ul

Circularization reaction

Exp.1 Exon capture experiment

   Sheared NA10835(~300ng/ul)        3ul
   E55kv1 probes 107nM              17ul
   10X Ampligase buffer           2.25ul

95C 10min -> 60C 40h -> add 2.5ul NSL mix -> 60C 1h -> (95C 2min -> 60C 1h) x 20 -> 37C 1min -> add 2ul exo I&III, 1ul T7 exo -> 37C 2h -> 95C 2min -> 4c hold