"
Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
From 2014.igem.org
(Difference between revisions)
| Line 5: | Line 5: | ||
{{Team:HokkaidoU_Japan/LargeDropDown/CSS}} | {{Team:HokkaidoU_Japan/LargeDropDown/CSS}} | ||
<html> | <html> | ||
| - | |||
<div id="background"> | <div id="background"> | ||
<!--begin header--> | <!--begin header--> | ||
| Line 13: | Line 12: | ||
<div id="header-bottom"> | <div id="header-bottom"> | ||
<div id="header-bottom-contents"> | <div id="header-bottom-contents"> | ||
| - | + | <div class="header-title"> | |
| + | <div class="header-title-top"> | ||
| + | Notebook | ||
| + | </div> | ||
| + | <div class="header-title-bottom"> | ||
| + | Lab Documents | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="header-picture"> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/4/44/HokkaidoU_Header_ki.png"> | ||
| + | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 14:49, 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!
