"
Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
From 2014.igem.org
(Difference between revisions)
Kakurehana (Talk | contribs) |
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 22:46, 17 October 2014
Notebook
Lab Documents
Stem Region Replacement
-
Start
-
PCR
pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ KOD FX NEO -
Transformation
R0010 on ice 30min each of 5.0µL DNA to DH5α 42℃ 1min on ice 2min -
Digestion
Cut R0010 with PstI, SpeI (using 10xH buffer) and pHN1257 with XbaI, PstI (using 10xM buffer) -
Ligation
Ligate R0010 with pHN1257 -
Transformation
Ligation products on ice 30min each of 5.0µL DNA to DH5α 42℃ 1min on ice 2min -
Colony PCR
R0010-stem-dT 100up_F pHN1257terminator_R -
mini-Prep
R0010-stem-dT -
PCR(confirm)
R0010-stem-dT 100up_F pHN1257terminator_R KOD FX NEO -
Complete!
