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Team:HokkaidoU Japan/Notebook/Length/as90 120
From 2014.igem.org
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| - | Cut as 90, as120, H-stem with NcoI, XhoI(using 10×Cut Smart) | + | Cut as 90, as120, H-stem-vector with NcoI, XhoI(using 10×Cut Smart) |
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| - | as90 | + | as90 on H-stem-vector XhoI_RBS_F Tm:68.2℃ as90_NcoI_R Tm:79.9℃ as120 on H-stem-vector XhoI_RBS_F Tm:68.2℃ as120_NcoI_R Tm:66.9℃ |
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Revision as of 15:08, 17 October 2014
Notebook
Lab Documents
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-vector 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 on H-stem-vector XhoI_RBS_F Tm:68.2℃ as90_NcoI_R Tm:79.9℃ as120 on H-stem-vector XhoI_RBS_F Tm:68.2℃ as120_NcoI_R Tm:66.9℃ -
Liquid Culture
as90, as120 on H-stem -
Mini-prep
as90, as120 on H-stem -
Dye terminator
as90, as120 on H-stem -
Complete!
