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===Validation of results=== *Successfully completely fragmented 6 ng or DNA, and partially fragmented 25 ng DNA. SDS-PAGE gel matches that which was provided by Gosta and Rickard. *After concentrating in amicon filters, A600 was 7.44 or 86.01 uM. Thus final concentration after added 1.05 volumes glycerol, .33 volumes 2X tn5 dialysis buffer (total of 2.38X dilution factor), is 36.13 uM in 5.84 mL. '''6/10/2014''' *with 25 ng Jurkat DNA (unconcentrated transposase) *annealed sc1-T7tspn-NoP-bot to T7tspn-top2(10/30) *tsp complex made by incubating at RT for ~30 mins *reaction composition 2 uL 5X custom tagmentation buffer (D6) 1 uL pure Jurkat DNA (25 ng) 1 uL tspsome 1 uL 2X Lysis Buffer 5 uL Nuclease free water _____________ 10 uL total [[File:ZhangLab 2 2014-06-10 20hr 02min-labeled.jpg|600px]] '''6/11/2014''' *with 6 ng DNA (unconcentrated transposase) *annealed sc1-T7tspn-NoP-bot to T7tspn-top2(10/30) *tsp complex made by incubating at RT for ~30 mins *reaction composition 1 uL 5X tagmetnation buffer 1 uL pure Jurkat DNA (6 ng) 1 or 2 uL transposome complex 1 uL 2X Lysis buffer 1 or 0 uL H2O [[File:ZhangLab 2 2014-06-11 12hr 59min-labeled.jpg|600px]] *Example of Tn5 band from gosta [[File:ZhangLab 2 2014-06-11 Gosta SDS-PAGE gel image.png|600px]] *original SDS-PAGE gel image [[File:ZhangLab 2 2014-06-11 SDS-PAGE gel tn5 original image.jpg|600px]] *SDS-PAGE gel with ladder labeled [[File:ZhangLab 2 2014-06-11 SDS-PAGE gel tn5.png|600px]]
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