"
Team:HokkaidoU Japan/Notebook/asB0034/asB0034-construction
From 2014.igem.org
(Difference between revisions)
(Created page with "{{Team:HokkaidoU_Japan/CSS}} {{Team:HokkaidoU_Japan/Notebook/CSS}} <html> <div class="wrapper"> <div id="hokkaidou-contents"> <div class="workflow-container"> <ul cla...") |
|||
| Line 4: | Line 4: | ||
<div class="wrapper"> | <div class="wrapper"> | ||
<div id="hokkaidou-contents"> | <div id="hokkaidou-contents"> | ||
| + | <h2>asB0034 Construction</h2> | ||
<div class="workflow-container"> | <div class="workflow-container"> | ||
<ul class="steps"> | <ul class="steps"> | ||
Revision as of 09:35, 25 September 2014
asB0034 Construction
-
Start
-
Primer Annealing
asB0034-f Tm: 68.7°C asB0034-r Tm: 68.7°C -
Digestion
Cut pHN1257 with NcoI, XhoI(using 10×cut smart) -
Gel extraction
digested pHN1257 -
Ethanol Precipetation
digested pHN1257 -
Ligation
Ligate pHN1257 with asB0034 -
Transformation
Ptrc-stem-asB0034-stem(pHN1257) on ice 30min each of 5.0µL DNA to DH5α 42°C 1min on ice 2min -
Colony PCR
Ptrc-stem-asB0034-stem(pHN1257) pHN1257_up_F Tm:60.1 asB0034-R Tm:68.7 -
Culture
Ptrc-stem-asB0034-stem -
mini-Prep
Ptrc-stem-asB0034-stem -
Transformation
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42C 30sec on ice 2min -
Complete!
