Difference between revisions of "Team:Bulgaria/IMPROVE"

 
(6 intermediate revisions by the same user not shown)
Line 261: Line 261:
 
 
 
p{
 
p{
−
text-align: center !important;
+
text-align: left !important;
−
margin-left: 128px;
+
margin-left: 64px;
−
margin-right: 128px;
+
margin-right: 64px;
 
font-family: 'Montserrat', sans-serif !important;
 
font-family: 'Montserrat', sans-serif !important;
 
font-size: 20px !important;
 
font-size: 20px !important;
Line 452: Line 452:
 
#myBtn:hover {
 
#myBtn:hover {
 
     background-color: #555; /* Add a dark-grey background on hover */
 
     background-color: #555; /* Add a dark-grey background on hover */
 +
}
 +
#downlaodlink{
 +
cursor: pointer;
 +
font-family: 'Wallpoet', sans-serif;
 +
}
 +
 +
#downlaodlink:hover{
 +
color: blue;
 +
}
 +
#n1{
 +
opacity: 1;
 +
background-image: url("https://static.igem.org/mediawiki/2018/8/81/T--Bulgaria--results-petrita.jpg");
 +
height: 450px;
 +
background-color: #0A1215;
 +
background-position: center;
 +
background-repeat: no-repeat;
 +
background-size: cover;
 
}
 
}
 
</style>
 
</style>
Line 519: Line 536:
 
<!-- CUSTOM TOP -->
 
<!-- CUSTOM TOP -->
 
<div class="contentHolder" id="topTitle">
 
<div class="contentHolder" id="topTitle">
−
<a id="pagetitle">PROJECT / IMPROVE</a>
+
<a id="pagetitle">IMPROVE</a>
 
</div>
 
</div>
 
</div>
 
</div>
Line 528: Line 545:
 
<!-- FOR NEW ONE COPY FROM HERE - CUSTOM CONTENT HOLDER -->
 
<!-- FOR NEW ONE COPY FROM HERE - CUSTOM CONTENT HOLDER -->
 
<div class="contentHolder" id="PROJECTIMPROVE">
 
<div class="contentHolder" id="PROJECTIMPROVE">
−
<h1>TITLE ONE</h1>
 
 
<p>
 
<p>
 
We improved the gRNA expression vector pSB1C3-gRNA - part BBa_K2515002
 
We improved the gRNA expression vector pSB1C3-gRNA - part BBa_K2515002
Line 540: Line 556:
 
and can be eliminated by cultivation at 42 o C. </p>
 
and can be eliminated by cultivation at 42 o C. </p>
 
<p>
 
<p>
−
Link: http://parts.igem.org/Part:BBa_K2515002
+
Link: <a id="downlaodlink" href="http://parts.igem.org/Part:BBa_K2515002">http://parts.igem.org/Part:BBa_K2515002</a>
 
<p>
 
<p>
  
Line 552: Line 568:
 
origin and can be used as a template in PCR amplification reactions. </p>
 
origin and can be used as a template in PCR amplification reactions. </p>
 
<p>  
 
<p>  
−
Link: http://parts.igem.org/Part:BBa_K2847002
+
Link: <a id="downlaodlink" href="http://parts.igem.org/Part:BBa_K2847002">http://parts.igem.org/Part:BBa_K2847002</a>
−
</p>
+
</p></div>
−
<p>
+
<!-- CUSTOM CONTENT HOLDER -->
−
https://static.igem.org/mediawiki/2018/8/81/T--Bulgaria--results-petrita.jpg
+
<div class="contentHolder" id="n1">
−
</p>
+
 
−
<p>
+
</div>
−
Fig. 1 pSB1C3-pSC101* T-sensitive origin. An illustration of our concept for a part
+
<!-- CUSTOM CONTENT HOLDER -->
 +
<div class="contentHolder" id="second">
 +
<div style="font-size: 15px; margin-left: 70px; margin-right: 70px;">
 +
<strong>Fig. 1 pSB1C3-pSC101* T-sensitive origin. </strong><br>An illustration of our concept for a part
 
improvement. The normal origin of replication found in pSB1C3 was substituted by the
 
improvement. The normal origin of replication found in pSB1C3 was substituted by the
 
pSC101* T-sensitive origin. The resulting vector was a low copy number plasmid and
 
pSC101* T-sensitive origin. The resulting vector was a low copy number plasmid and
 
required decreased antibiotic levels for growth (left). If needed, it could be successfully
 
required decreased antibiotic levels for growth (left). If needed, it could be successfully
−
cured by an overnight cultivation at 42 o C (right).
+
cured by an overnight cultivation at 42°C (right).
−
 
+
−
</p>
+
 
</div>
 
</div>
 +
</div>
 +
 
<!-- TO HERE - CUSTOM CONTENT HOLDER -->
 
<!-- TO HERE - CUSTOM CONTENT HOLDER -->
  
Line 631: Line 650:
 
       <a href="https://2018.igem.org/Team:Bulgaria/MEDALSILVER">SILVER</a>
 
       <a href="https://2018.igem.org/Team:Bulgaria/MEDALSILVER">SILVER</a>
 
  <a href="https://2018.igem.org/Team:Bulgaria/MEDALGOLD">GOLD</a>
 
  <a href="https://2018.igem.org/Team:Bulgaria/MEDALGOLD">GOLD</a>
 +
<a href="https://2018.igem.org/Team:Bulgaria/MEDALAWARD">SPECIAL AWARD</a>
 
</div>
 
</div>
 
   </div>  
 
   </div>  

Latest revision as of 23:28, 17 October 2018

PROJECT USE IS IT SAFE USED ORGANISM STRAINS POTENTIAL RISKS WASTE TREATMENT EMERGENCY REACTIONS PROTECTIVE EQUIPMENT OTHER RULES

We improved the gRNA expression vector pSB1C3-gRNA - part BBa_K2515002 from the Registry. It was originally designed by the iGEM Bulgaria 2017 team for easy cloning and expression of gRNAs in E. coli. Nevertheless, it is a high copy number vector with no easy option for a plasmid curing. To overcome these limitations, we PCR amplified the pSC101-based thermosensitive origin of replication from the pE-FLP vector (Plasmid #45978). Next, we developed an aqua cloning-based approach to substitute the original pSB1C3 origin with this new part. The resulting vector was low copy number (that is sufficient for gRNA expression due to the high efficiencies of the CRISPR/Cas9 systems) and can be eliminated by cultivation at 42 o C.

Link: http://parts.igem.org/Part:BBa_K2515002

NB! The generated vector is a low copy number plasmid, thus, one needs to use reduced antibiotic concentrations and the colony growth requires more time. In addition, the pSC101 ori contains a SpeI restriction site and, therefore, is not BioBrick RCF10- compatible! If you need to use it – adopt the aqua cloning procedure from the part’s page in the Registry.

We submitted a new part BBa_K2847002 – it contains the thermosensitive replication origin and can be used as a template in PCR amplification reactions.

Link: http://parts.igem.org/Part:BBa_K2847002

Fig. 1 pSB1C3-pSC101* T-sensitive origin.
An illustration of our concept for a part improvement. The normal origin of replication found in pSB1C3 was substituted by the pSC101* T-sensitive origin. The resulting vector was a low copy number plasmid and required decreased antibiotic levels for growth (left). If needed, it could be successfully cured by an overnight cultivation at 42°C (right).