"
Team:HokkaidoU Japan/Notebook/asB0034/asB0034-construction
From 2014.igem.org
(Difference between revisions)
Kakurehana (Talk | contribs) |
|||
| Line 5: | Line 5: | ||
{{Team:HokkaidoU_Japan/LargeDropDown/CSS}} | {{Team:HokkaidoU_Japan/LargeDropDown/CSS}} | ||
<html> | <html> | ||
| + | <link href='http://fonts.googleapis.com/css?family=Merriweather' rel='stylesheet' type='text/css'> | ||
<div id="background"> | <div id="background"> | ||
<!--begin header--> | <!--begin header--> | ||
Revision as of 22:55, 17 October 2014
Notebook
Lab Documents
asB0034 Construction
-
Start
-
Primer Annealing
asB0034_F Tm: 68.7℃ asB0034_R Tm: 68.7℃ -
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℃ 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 42℃ 30sec on ice 2min -
Complete!
