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Team:HokkaidoU Japan/Notebook/Length/as90 120
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Revision as of 11:40, 12 October 2014
Anti-sense 90, 120 Construction
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Start
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PCR & PCR purification
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R0040-B0034-E1010-B0015 as RBS XhoI, 90bp as mRFP Nco as RBS XhoI, 120bp as mRFP Nco
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Digestion
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Cut as 90, as120, H-stem with NcoI, XhoI(using 10×Cut Smart)
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Ethanol precipitation
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as90, as120
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Gel Extraction
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H-stem
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Dephophorylation
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H-stem
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Ligation
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Ligate as90, as120 with H-stem
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Transformation
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as90, as120 on H-stem 5µL DNA to DH5α Turbo
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Colony PCR
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as90, as120 on H-stem
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Liquid Culture
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as90, as120 on H-stem
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Mini-prep
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as90, as120 on H-stem
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Dye terminator
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as90, as120 on H-stem
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Complete!
