"
Team:HokkaidoU Japan/Notebook/Pre experiment/Stem replacement
From 2014.igem.org
(Difference between revisions)
| (7 intermediate revisions not shown) | |||
| 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--> | ||
| Line 13: | Line 14: | ||
<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> | ||
| Line 29: | Line 40: | ||
<div class="ldd_submenu" style="left:-415px; width:970px;"> | <div class="ldd_submenu" style="left:-415px; width:970px;"> | ||
<ul style="margin-left:20px;"> | <ul style="margin-left:20px;"> | ||
| - | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/ | + | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Overview</a></li> |
| - | + | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Background">Background</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Overview/Achievements">Achievements</a></li> | ||
| Line 51: | Line 61: | ||
<ul> | <ul> | ||
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Length Variation</a></li> | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Length Variation</a></li> | ||
| - | <li class="ldd_contents"><a href="">Overview</a></li> | + | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length">Overview</a></li> |
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Method">Method</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Method">Method</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Results">Results</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Projects/Length#Results">Results</a></li> | ||
| Line 73: | Line 83: | ||
</ul> | </ul> | ||
<ul> | <ul> | ||
| - | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey">High | + | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey">High School</a></li> |
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Education">Education</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Education">Education</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Survey">Survey</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Survey#Survey">Survey</a></li> | ||
| Line 80: | Line 90: | ||
<li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion">Discussion</a></li> | <li class="ldd_heading"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion">Discussion</a></li> | ||
<li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Background">Background</a></li> | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Background">Background</a></li> | ||
| - | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion# | + | <li class="ldd_contents"><a href="https://2014.igem.org/Team:HokkaidoU_Japan/Outreach/Discussion#Evaluation">Evaluation</a></li> |
</ul> | </ul> | ||
</div> | </div> | ||
| Line 131: | Line 141: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ KOD FX NEO | + | pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃ |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD FX NEO | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 152: | Line 165: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010 | + | R0010 |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | each of 5.0µL DNA to DH5α | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 173: | Line 189: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Cut R0010 with PstI, SpeI (using 10xH buffer) | + | Cut R0010 with PstI, SpeI (using 10xH buffer) |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | Cut pHN1257 with XbaI, PstI (using 10xM buffer) | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 215: | Line 234: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ligation products | + | Ligation products |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | each of 5.0µL DNA to DH5α | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 236: | Line 258: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem- | + | R0010-stem-B0015 with 100up_F pHN1257terminator_R |
</div> | </div> | ||
</div> | </div> | ||
| Line 244: | Line 266: | ||
<div class="step-tile"> | <div class="step-tile"> | ||
<h2 class="step-title"> | <h2 class="step-title"> | ||
| - | + | Mini-Prep | |
</h2> | </h2> | ||
</div> | </div> | ||
| Line 257: | Line 279: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem- | + | R0010-stem-B0015 |
</div> | </div> | ||
</div> | </div> | ||
| Line 278: | Line 300: | ||
</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | R0010-stem-dT 100up_F pHN1257terminator_R KOD FX NEO | + | R0010-stem-dT with 100up_F pHN1257terminator_R |
| + | </div> | ||
| + | <div class="step-recipe"> | ||
| + | KOD FX NEO | ||
</div> | </div> | ||
</div> | </div> | ||
| Line 297: | Line 322: | ||
<div id="footer-content"> | <div id="footer-content"> | ||
<div id="footer-logo"> | <div id="footer-logo"> | ||
| - | <a href="http://igemhokkaidou.com | + | <a href="http://igemhokkaidou.wordpress.com"><img style="height:150px;position:relative;" src="https://static.igem.org/mediawiki/2014/3/39/HokkaidoU_logo_transparent.png"></a> |
</div> | </div> | ||
<div id="footer-twitter"> | <div id="footer-twitter"> | ||
Latest revision as of 15:26, 9 September 2015
Notebook
Lab Documents
Stem Region Replacement
-
Start
-
PCR
pHN1257 stem_F Tm:70.6℃ terminator_R Tm:71.4℃KOD FX NEO -
Transformation
R0010each of 5.0µL DNA to DH5α -
Digestion
Cut R0010 with PstI, SpeI (using 10xH buffer)Cut pHN1257 with XbaI, PstI (using 10xM buffer) -
Ligation
Ligate R0010 with pHN1257 -
Transformation
Ligation productseach of 5.0µL DNA to DH5α -
Colony PCR
R0010-stem-B0015 with 100up_F pHN1257terminator_R -
Mini-Prep
R0010-stem-B0015 -
PCR(confirm)
R0010-stem-dT with 100up_F pHN1257terminator_RKOD FX NEO -
Complete!
