Team:EPF-Lausanne/Protocols/Klenow

From 2009.igem.org

(Difference between revisions)
(New page: Klenow protocol New protocol used with no purification step, everything is prepared to go trough all reaction up to the end of the digestion, ready for the ligation without going through a...)
Line 1: Line 1:
-
Klenow protocol
+
<html>
-
New protocol used with no purification step, everything is prepared to go trough all reaction up to the end of the digestion, ready for the ligation without going through any step of purification. This will avoid losing DNA due to the small size of the TrpO (127bp).
+
<head>
-
Goal: we want 100ng/ul od DNA at the end
+
-
length of TrpO:127bp
+
-
molecular weight: 78,522 . 10^3 g/mol
+
-
→ 100ng/ul in 50ul → 6,367.10^-11 mol of TrpO a the end, that's to say 6,367.10^-11 of each primer
+
-
Trp Operon-Rev: 1,560.10^3M → 2.56 ul Trp Operon-Fwd: 1,560.10^3M → 2.56 ul
+
-
first make a dilution at 25ul in 500ul → 8ul in 492ul of MQ
+
-
1. Klenow
 
-
dNTPs final concentr = 1mM
 
-
3.64ul of NEB2
 
-
0.37ul of BSA 100x
 
-
2.56ul of each primer at 25uM
 
-
32.38ul of MQ
 
-
→ final: 36.4ul
 
-
Thermal cycler:
+
<style>
-
94°c for 5min
+
/*prop. de la premiere ligne*/
-
0.1°C/s to 74°C for 5 min
+
.firstHeading {
-
0.1°C/s to 37°C
+
  display:none;
-
Annealing temperature: 79.1°C (due to some website)
+
}
 +
.top-section {
 +
  display:none;
 +
}
-
1.' 1ul Klenow + 1.6ul dNTPs = vol 39ul Incubate 1h30 at 37°C, then inactivate 20 min at 75°C : 0.1°C/s to 37°C
+
/*box igem en bas*/
 +
#footer-box {
 +
  float: bottom;
 +
}
-
2. Digestion
+
/*couleur du box ou on peut printer*/
-
Add 0.5 ul of each enzyme (EcoRI - HF / NheI) → vol: 40.0ul Incubate 2h at 37°C, then inactivate 20min at 80°C. Then cool down at 37°C (0.1°C/s)
+
#content {
 +
          /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg") fixed;*/
 +
background-color:#CEE0EC;
 +
          border-width: 0px;
 +
}
-
3. Dephosphorylation
+
/*couleur du sous-box a l'intérieur de content, garder la couleur a neutre*/
-
Add 5ul of antartic phosphatase buffer + 5ul of phosphatase enzyme → total volume: 50ul Incubate 2h at 37°C, then inactivate 10min at 65°C. Then cool down slowly (0.1°C/s)
+
#bodyContent {
-
1ul is needed to dephosphorylate 1-5mg of vector pUC19
+
          padding: 0px;
 +
          margin: 0px;
 +
 
 +
}
 +
/*------------------------calendar-----------------------*/
 +
 
 +
.heading {
 +
background-color: #007CBC;
 +
font-size: 20px;
 +
color: white;
 +
}
 +
.dow {
 +
background-color: #E5E9FF;
 +
font-size: 16px;
 +
color: #007CBC;
 +
}
 +
.month {
 +
border: 1px;
 +
}
 +
 
 +
/*couleur de la page contenant le wiki*/
 +
body {
 +
          /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg") fixed;*/
 +
background-color:#CEE0EC;
 +
}
 +
 
 +
.epfl09 {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 50px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
 
 +
.epfl09team {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg");*/
 +
        background-image:url('https://static.igem.org/mediawiki/2009/c/cd/P6179_back.jpg');
 +
        background-repeat:no-repeat;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 50px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
 
 +
.epfl09model {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg");*/
 +
        background-image:url('https://static.igem.org/mediawiki/2009/f/f9/Mod_back.jpg');
 +
        background-repeat:no-repeat;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 50px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
 
 +
.epfl09proj {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg");*/
 +
        background-image:url('https://static.igem.org/mediawiki/2009/6/69/Proj_back.png');
 +
        background-repeat:no-repeat;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 50px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
 
 +
.epfl09lab {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg");*/
 +
        background-image:url('https://static.igem.org/mediawiki/2009/f/f2/Lab_back.jpg');
 +
        background-repeat:no-repeat;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 50px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
.epfl09hum {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg");*/
 +
        background-image:url('https://static.igem.org/mediawiki/2009/f/f3/IMG_0138_back.jpg');
 +
        background-repeat:no-repeat;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 50px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
 
 +
.epfl09bis {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  804px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 0px #007cbc;
 +
        padding-top: 10px;
 +
        padding-left: 120px;
 +
        padding-right: 5px;
 +
    }
 +
 
 +
.epfl09front {
 +
position: relative;
 +
        min-height:400px;
 +
        width:  929px;
 +
        left: 40px;
 +
        top: -155px;
 +
        bottom: 0px;
 +
        right: 0px;
 +
        z-index:0;
 +
        background-color: white;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
        border-top: solid 1px #007cbc;
 +
        padding-top: 15px;
 +
        padding-left: 0px;
 +
        padding-right: 0px;
 +
    }
 +
 
 +
.epfl09bouchon {
 +
position: absolute;
 +
        float: bottom;
 +
        width:  929px;
 +
        left: 45px;
 +
        bottom: 0px;
 +
        z-index:-1;
 +
        min-height: 180px;
 +
        background-color: white;
 +
        border-left: solid 1px #007cbc;
 +
        border-right: solid 1px #007cbc;
 +
    }
 +
 
 +
.epfltrick {
 +
position: absolute;
 +
        top: 220px;
 +
        left: 0px;
 +
        right: 830px;
 +
        bottom: 0px;
 +
        z-index:1;
 +
    }
 +
 
 +
 
 +
 
 +
/********************************************************************************************************/
 +
/****                              css pr menu                                                      ****/
 +
/********************************************************************************************************/
 +
 
 +
 
 +
.wrapper1{
 +
color: #44433f;
 +
font: 14px "Futura Medium", "Myriad Pro", "Gill Sans", Helvetica, Verdana, Arial, sans-serif;
 +
margin: 0;
 +
padding: 0px 0 0;
 +
}
 +
.wrapper1 a{
 +
color: #E5F2FB;
 +
text-decoration: none;
 +
}
 +
.wrapper1 a:hover {
 +
color: #09548B;
 +
}
 +
.wrapper1 p {
 +
margin: 0 0 17px;
 +
padding: 0;
 +
line-height: 18px;
 +
}
 +
.wrapper {
 +
/*width: 710px;*/
 +
margin: 20px auto;
 +
}
 +
.nav {
 +
background: url(https://static.igem.org/mediawiki/2009/4/43/Nav_bg.png) repeat-x;
 +
float: left;
 +
}
 +
.nev-wrapper {
 +
clear: both;
 +
float: left;
 +
}
 +
.nav-left {
 +
background: url(https://static.igem.org/mediawiki/2009/d/d0/Nav_left.png) no-repeat top left;
 +
float: left;
 +
width: 11px;
 +
height: 41px;
 +
}
 +
.nav-right {
 +
background: url(https://static.igem.org/mediawiki/2009/0/0f/Nav_right.png) no-repeat top right;
 +
float: left;
 +
width: 11px;
 +
height: 41px;
 +
}
 +
.nav ul {
 +
/*width: 648px;*/
 +
height: 38px;
 +
float: left;
 +
margin: 0;
 +
padding-top: 3px;
 +
list-style: none;
 +
font-size: 15px;
 +
}
 +
.nav li {
 +
float: left;
 +
padding: 0 7px;
 +
background: url(https://static.igem.org/mediawiki/2009/4/41/Split.png) no-repeat right center;
 +
position: relative;
 +
z-index: 1;
 +
}
 +
.nav li.last {
 +
background:none;
 +
}
 +
.nav li:hover {
 +
z-index:2;
 +
}
 +
.nav li a {
 +
display: block;
 +
line-height: 38px;
 +
overflow: hidden;
 +
float: left;
 +
}
 +
a .menu-left {
 +
background: url(https://static.igem.org/mediawiki/2009/5/55/Menu_left.gif) no-repeat left top;
 +
width: 8px;
 +
height: 32px;
 +
line-height: 35px;
 +
display: block;
 +
float: left;
 +
}
 +
a .menu-mid {
 +
background: url(https://static.igem.org/mediawiki/2009/a/ac/Menu_mid.gif) repeat-x top left;
 +
height: 32px;
 +
line-height: 35px;
 +
display: block;
 +
float: left;
 +
}
 +
a .menu-right {
 +
background: url(https://static.igem.org/mediawiki/2009/f/f9/Menu_right.gif) no-repeat top left;
 +
width: 8px;
 +
height: 32px;
 +
line-height: 35px;
 +
display: block;
 +
float: left;
 +
}
 +
.nav li a:hover .menu-left,
 +
.nav li.active a .menu-left,
 +
.nav li:hover a .menu-left,
 +
.nav li a:hover .menu-mid,
 +
.nav li.active a .menu-mid,
 +
.nav li:hover a .menu-mid,
 +
.nav li a:hover .menu-right,
 +
.nav li.active a .menu-right,
 +
.nav li:hover a .menu-right {
 +
background-position: 0 -37px;
 +
line-height: 35px;
 +
}
 +
.nav li a:hover,
 +
.nav li.active a,
 +
.nav li.hover a,
 +
.nav li:hover a {
 +
color: #09548B;
 +
}
 +
.nav li:hover .sub,
 +
.nav li.hover .sub {
 +
display:block;
 +
}
 +
.nav li .sub {
 +
display: none;
 +
position: absolute;
 +
top: 27px;
 +
left: 6px;
 +
background: url(https://static.igem.org/mediawiki/2009/2/2a/Submenu_top.png) no-repeat;
 +
width: 186px;
 +
padding-top: 9px;
 +
}
 +
.nav li ul {
 +
background: url(https://static.igem.org/mediawiki/2009/e/eb/Submenu_bg.png) repeat-y;
 +
width: 162px;
 +
height: auto;
 +
margin: 0;
 +
padding: 0 12px 10px;
 +
list-style: none;
 +
font-size: 14px;
 +
}
 +
 
 +
.nav li:hover li,
 +
.nav li.active li {
 +
width: 100%;
 +
padding: 1px 0 2px;
 +
border-bottom: 1px #C1D9F0 dashed;
 +
background: none !important;
 +
}
 +
.nav li:hover li a,
 +
.nav li.active li a {
 +
color: #09548B;
 +
background: none !important;
 +
line-height: normal;
 +
width: 156px;
 +
padding: 8px 3px 3px;
 +
text-indent: 1px;
 +
}
 +
.nav li:hover li a:hover,
 +
.nav li.active li a:hover {
 +
color: #fff;
 +
background: #165B9F !important;
 +
text-decoration: none;
 +
line-height: normal;
 +
}
 +
/*IE*/
 +
.nav li li a:hover,
 +
.nav li li a:hover {
 +
color: #fff;
 +
background: #165B9F !important;
 +
text-decoration: none;
 +
line-height: normal;
 +
}
 +
/**/
 +
.nav .btm-bg {
 +
background: url(https://static.igem.org/mediawiki/2009/2/2d/Submenu_bottom.png) no-repeat;
 +
width: 205px;
 +
height: 9px;
 +
overflow: hidden;
 +
clear: both;
 +
}
 +
.content {
 +
width: 670px;
 +
background: transparent url(https://static.igem.org/mediawiki/2009/1/1e/Content_bg.png) repeat-y;
 +
float: left;
 +
padding: 10px 20px;
 +
}
 +
.content h1 {
 +
color: #333;
 +
font-weight: 400;
 +
text-transform: uppercase;
 +
font-size: 18px;
 +
border-bottom: 1px dashed #C1D9F0;
 +
}
 +
.content h2 {
 +
font-weight: 400;
 +
text-transform: uppercase;
 +
font-size: 14px;
 +
padding-left: 10px;
 +
margin-bottom: -5px;
 +
}
 +
.content p {
 +
padding: 0 15px;
 +
text-align: justify;
 +
}
 +
.content-bottom {
 +
width: 710px;
 +
background: transparent url(https://static.igem.org/mediawiki/2009/0/01/Content_bottom.png) no-repeat;
 +
height: 13px;
 +
float: left;
 +
}
 +
</style>
 +
 
 +
<style type="text/css">
 +
<!--
 +
.demo  {color:#000033; background-color:#cccccc; layer-background-color:#cccccc;
 +
        position:absolute; top:100px; left:100px; width:480px; height:280px;
 +
        z-index:99;  visibility:hidden;}
 +
-->
 +
</style>
 +
 
 +
 
 +
 
 +
</head>
 +
 
 +
<div style="position: relative; left: 20px; top: 0px; right: 0px; height: 140px; width: 284px; padding: 0px; margin: 0px; border: 0px solid white; z-index:1;">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne"><img border="0" src="https://static.igem.org/mediawiki/2009/5/51/Banniere2.png"></a>
 +
</div>
 +
 
 +
<div style="position: relative; left: 190px; top: -134px; padding: 0px; margin: 0px; border: 0px solid white;z-index:1;">
 +
 
 +
<div class="wrapper1">
 +
<div class="wrapper"  style="width:650px;">
 +
<div class="nav-wrapper">
 +
<div class="nav-left"></div>
 +
<div class="nav">
 +
<ul id="navigation" style="width:600px";>
 +
 
 +
<li class="#"><a href=https://2009.igem.org/Team:EPF-Lausanne target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Home</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
 
 +
 
 +
 
 +
 +
<li class="#">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Team" target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Team</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
<div class="sub">
 +
<ul>
 +
 
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Team" target="_self">Presentation</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Team#Where_we_are_from" target="_self">School</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Team_Management" target="_self">Team Management</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Brainstorming" target="_self">Brainstorming</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Sponsors" target="_self">Sponsors</a>
 +
</li>
 +
</ul>
 +
<div class="btm-bg">
 +
</div></div>
 +
</li>
 +
 
 +
 
 +
<li class="#">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Project_Abstract" target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Project</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
<div class="sub">
 +
<ul>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Project_Abstract" target="_self">Project Overview</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/LOVTAP" target="_self">Introduction LovTAP</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Strategy" target="_self">Strategy</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Experimental_Setup" target="_self">Experimental Setup</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/LOVTAP_Results" target="_self">Results</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Parts" target="_self">Parts submitted to the Registry</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Future_directions" target="_self">Future directions</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/References" target="_self">References</a>
 +
</li>
 +
</ul>
 +
 
 +
<div class="btm-bg">
 +
</div></div>
 +
</li>
 +
 
 +
 
 +
 
 +
<li class="#">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Modeling_overview" target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Modeling</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
<div class="sub">
 +
<ul>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Modeling_overview" target="_self">Modeling overview</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Analysis methods" target="_self">Analysis Methods</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Results" target="_self">Results</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Information_&_references" target="_self">References</a>
 +
</li>
 +
</ul>
 +
 
 +
<div class="btm-bg">
 +
</div></div>
 +
</li>
 +
 
 +
 
 +
<li class="#">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Social_overview" target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Human practices</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
 
 +
<div class="sub">
 +
<ul>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Social_overview" target="_self">Overview</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Survey" target="_self">Survey</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Talks" target="_self">Talks</a>
 +
</li>
 +
<li>
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Media" target="_self">Media</a>
 +
</li>
 +
</ul>
 +
 
 +
<div class="btm-bg">
 +
</div></div>
 +
</li>
 +
 
 +
<li class="#">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Safety" target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Safety</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
 
 +
<li class="#">
 +
<a href="https://2009.igem.org/Team:EPF-Lausanne/Notebook" target="_self">
 +
<span class="menu-left"></span>
 +
<span class="menu-mid">Lab</span>
 +
<span class="menu-right"></span>
 +
</a>
 +
 
 +
<div class="sub">
 +
<ul>
 +
 
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Last_News" target="_self">Last News</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Notebook" target="_self">Notebook</a>
 +
</li>
 +
<li>
 +
<a href= "https://2009.igem.org/Team:EPF-Lausanne/Protocols" target="_self">Protocols used</a>
 +
</li>
 +
</ul>
 +
<div class="btm-bg">
 +
</div></div>
 +
</li>
 +
 
 +
 
 +
</div>
 +
<div class="nav-right"></div>
 +
<br><br>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
 
 +
 
 +
</html>
 +
<html>
 +
<script type="text/javascript" language="JavaScript"><!--
 +
function HideContent(d) {
 +
document.getElementById(d).style.display = "none";
 +
}
 +
function ShowContent(d) {
 +
document.getElementById(d).style.display = "block";
 +
}
 +
function ReverseDisplay(d) {
 +
if(document.getElementById(d).style.display == "none") { document.getElementById(d).style.display = "block"; }
 +
else { document.getElementById(d).style.display = "none"; }
 +
}
 +
//--></script></html>

Revision as of 19:06, 20 October 2009



Retrieved from "http://2009.igem.org/Team:EPF-Lausanne/Protocols/Klenow"