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Team:HokkaidoU Japan/Notebook/Length/as90 120
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Anti-sense 90, 120 Construction | Anti-sense 90, 120 Construction | ||
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Revision as of 11:51, 12 October 2014
Anti-sense 90, 120 Construction
-
Start
-
PCR & PCR purification
R0040-B0034-E1010-B0015 as RBS XhoI, 90bp as mRFP Nco as RBS XhoI, 120bp as mRFP Nco -
Digestion
Cut as 90, as120, H-stem with NcoI, XhoI(using 10×Cut Smart) -
Ethanol precipitation
as90, as120 -
Gel Extraction
H-stem -
Dephophorylation
H-stem -
Ligation
Ligate as90, as120 with H-stem -
Transformation
as90, as120 on H-stem 5µL DNA to DH5α Turbo -
Colony PCR
as90, as120 on H-stem -
Liquid Culture
as90, as120 on H-stem -
Mini-prep
as90, as120 on H-stem -
Dye terminator
as90, as120 on H-stem -
Complete!
