Haynes Lab:Notebook/Protein-DNA fusions in living microbes/2013/11/21: Difference between revisions
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==Entry title== | ==Entry title== | ||
testing use of NH4SO4 to halt topoisomerase reaction. | |||
pSB1C3 250ng 250ng 250ng | |||
7.35uL 7.35uL 7.35uL | |||
topo eq. 0.5uL 0.5uL 0.5uL | |||
stop mechanism 4M am.sulfamate 4M am.acetate 0M ctrl | |||
16M stock: 15uL 15uL 0 | |||
NEB 4 3uL 3uL 3uL | |||
ddH20 up to 30 | |||
total 30uL 30uL 30uL | |||
incubation:37C 10min 10 10 | |||
ammonium acetate: 12.32g/10mL | |||
ammonium sulfamate: 18.26g/10mL | |||
stop mechanism (ammonium sulfamate and ammonium acetate) added before incubation. | |||
incubate at 37 for 10 minutes | |||
place in -20 C | |||
run on gel | |||
Revision as of 00:05, 3 December 2013
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Entry titletesting use of NH4SO4 to halt topoisomerase reaction. pSB1C3 250ng 250ng 250ng 7.35uL 7.35uL 7.35uL topo eq. 0.5uL 0.5uL 0.5uL stop mechanism 4M am.sulfamate 4M am.acetate 0M ctrl 16M stock: 15uL 15uL 0 NEB 4 3uL 3uL 3uL ddH20 up to 30 total 30uL 30uL 30uL incubation:37C 10min 10 10 ammonium acetate: 12.32g/10mL ammonium sulfamate: 18.26g/10mL stop mechanism (ammonium sulfamate and ammonium acetate) added before incubation. incubate at 37 for 10 minutes place in -20 C run on gel
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