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==Background== *Yesterday, I tried adjusting the Tn5 reaction volume to 15ul (see <http://genome-tech.ucsd.edu/LabNotes/index.php/Chris:LabNotes/SeaMicrobiome/Calendar/2015/2015-4-22>). However, only reactions 5&6 showed good smears (with rxn's 7&8 being very dim). This means that the 1ul 0.25x supermutant Tn5059 was sufficient for digesting 1ng of DNA but when we increased that to 2ul 0.25x Tn5059, there was over-digestion. Consequently, it looks like for the ~400pg DNA template input, I likewise over-digested the template leading to no product on the gel. *Today, I'll be working on another test, this time trying out different enzyme concentration inputs in order to lower the concentration to get some product. *In order to not run out of template, I'll be reverting back to the Catalina Harbor samples (particularly testing with CH-S-A, which was 63.2ng/ul). Consequently, I'll be running the following test: rxn Sample Template Input Enzyme Conc (1ul added) 1 CH-S-A 10ul 38pg/ul 0.25x 2 CH-S-A 10ul 38pg/ul 0.25x * 1/2d 3 CH-S-A 10ul 38pg/ul 0.25x * 1/4d 4 CH-S-A 10ul 38pg/ul 0.25x * 1/8d 5 CH-S-A 10ul 38pg/ul 0.25x * 1/16d 6 CH-S-A 1ul 1ng/ul 0.25x <- Pos Control 7 CH-S-A 1ul 1ng/ul 2ul 0.25x <- Verify results from previous test (rxn's 7/8 on 4/22) 8 NTC -- 0.25x <- NTC
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