Kun:LabNotes/ExonomeSeq/2008-1-9

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Revision as of 18:25, 9 January 2008 by KunZhang (talk | contribs) (New page: ==Exp.1 Sequencing library construction for the E55kv1/GM10835 amplicon 450bp band== Just realized that I should do Mme I digestion prior to other enzymatic reactions. ===Mme I digestion o...)
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Exp.1 Sequencing library construction for the E55kv1/GM10835 amplicon 450bp band

Just realized that I should do Mme I digestion prior to other enzymatic reactions.

Mme I digestion of amplicons

         dNTP ampilcon        dUTP amplicon          dUTP USER digestion
 DNA          20ul                 10ul                    15ul
 NEBuffer 4    3ul                  3ul                     3ul
 SAM(1mM)    1.5ul                1.5ul                   1.5ul
 Mme I         2ul                  2ul                     2ul
 H2O         3.5ul               13.5ul                   8.5ul

37C 1h -> 80C 20min -> 4C hold