Kun:LabNotes/ExonomeSeq/2008-1-9: Difference between revisions

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37C 1h -> 80C 20min -> 4C hold
37C 1h -> 80C 20min -> 4C hold
   [[Image:2008-01-09-Lib-construction.png|500px]]
   [[Image:2008-01-09-Lib-construction.png|500px]]
Digestion was not complete. I added 2ul more enzyme, 37C 2h -> 80C 20min.

Revision as of 01:39, 10 January 2008

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

 File:2008-01-09-Lib-construction.png

Digestion was not complete. I added 2ul more enzyme, 37C 2h -> 80C 20min.