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.