"
Team:Bordeaux/NotebookAugust12
From 2014.igem.org
(Difference between revisions)
| Line 25: | Line 25: | ||
<li>Insert: 20 ng/μl</li></ul> | <li>Insert: 20 ng/μl</li></ul> | ||
| - | <ul class="day-detail"> | + | <ul class="day-detail"><br><br> |
<li>Ligation<br> | <li>Ligation<br> | ||
<div style="margin-align:45px;"> | <div style="margin-align:45px;"> | ||
| Line 40: | Line 40: | ||
H2O qsp 10μl<br> | H2O qsp 10μl<br> | ||
</li> | </li> | ||
| - | + | <br><br> | |
<li>Making competent cells<br> | <li>Making competent cells<br> | ||
BL21 in LB+Amp plates</li> | BL21 in LB+Amp plates</li> | ||
Latest revision as of 01:34, 18 October 2014
12th August
Molecular biology
- Digestion
We want a DNA concentration of 100ng/μl
Time-Saver protocolDNA 10μl=> 15min at 37°C
Xba1 0,5μl
Spe1 0,5μl
Buffer 10X 5μl
H2O qsp 50μl
=> Inactivation of enzyme 20min at 80°C - Plasmid: 18 ng/μl
- Insert: 20 ng/μl
Final concentration after digestion :
- Ligation
ELP – 369bpPlasmid 4μl
SLP – 369bp
RLP – 357bp
Plasmid – 2270bp
Insert 1,8μl
Buffer 10X 1μl
Taq ligase 0,5μl
H2O qsp 10μl
- Making competent cells
BL21 in LB+Amp plates
