Difference between revisions of "Part:BBa K4887005"
 
Line 12: Line 12:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4887005 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4887005 SequenceAndFeatures</partinfo>
 +
 +
<html>
 +
<style>
 +
li {
 +
    list-style-type: none;
 +
    padding: 0;
 +
  }
 +
.bild{
 +
    max-width: 70%;
 +
    max-height: auto;
 +
}
 +
.red{
 +
    color:red;
 +
}
 +
.center {
 +
display: flex;
 +
justify-content: center;
 +
align-items: center;
 +
flex-direction: column;
 +
height: 50vh; /*注: 通过更改该数值可以减少 图片及配文 和 图片下面内容的间距 但是不能太小（>40）*/
 +
}
 +
 +
.unterschrift {
 +
text-align: center;
 +
}
 +
.toobig{
 +
max-width:50%;
 +
}
 +
 +
</style>
 +
 +
<h1>5.5 Results:</h1>
 +
<h2>(1) PCR detection for gene Cas9. </h2>
 +
Genome DNA of the transgenic lines was extracted from leaves of these two regenerated seedlings. PCR detection on the genomes was performed by using primers designed based on the Cas9 protein gene. As the result, the transgenic lines 23216004 and 23216005 were further validated <b class="red">(Fig. 1)</b>.
 +
<br />
 +
The sequences of the primers were as bellow: <br />
 +
<ul>
 +
<li>CAS9-F: 5’-atggactataaggaccacgacgg-3’</li>
 +
<li>CAS9-R: 5’-ttgtcgcctcccagctgagacag-3’</li>
 +
</ul>
 +
<div class="center">
 +
<img class="bild toobig" src="https://static.igem.wiki/teams/4887/wiki/images/images/part-detection-results-of-pcrs-for-gene-cas9.png"/>
 +
<div class="unterschrift">
 +
<b>
 +
Fig. 1 Detection results of PCR for gene Cas9.
 +
</b>
 +
</div>
 +
</div>
 +
</html>
  
  

Latest revision as of 00:48, 11 October 2023


Cas9

It is the gene of Cas9 (CRISPR-associated nuclease 9), a dual RNA-guided DNA endonuclease enzyme associated with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) adaptive immune system in Streptococcus pyogenes.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 907
    Illegal PstI site found at 2329
    Illegal PstI site found at 2533
    Illegal PstI site found at 2563
    Illegal PstI site found at 3775
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 907
    Illegal PstI site found at 2329
    Illegal PstI site found at 2533
    Illegal PstI site found at 2563
    Illegal PstI site found at 3775
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 368
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 907
    Illegal PstI site found at 2329
    Illegal PstI site found at 2533
    Illegal PstI site found at 2563
    Illegal PstI site found at 3775
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 907
    Illegal PstI site found at 2329
    Illegal PstI site found at 2533
    Illegal PstI site found at 2563
    Illegal PstI site found at 3775
    Illegal NgoMIV site found at 1195
    Illegal NgoMIV site found at 2299
    Illegal NgoMIV site found at 2372
    Illegal NgoMIV site found at 2857
    Illegal NgoMIV site found at 3766
    Illegal NgoMIV site found at 4227
    Illegal NgoMIV site found at 4246
  • 1000
    COMPATIBLE WITH RFC[1000]

5.5 Results:

(1) PCR detection for gene Cas9.

Genome DNA of the transgenic lines was extracted from leaves of these two regenerated seedlings. PCR detection on the genomes was performed by using primers designed based on the Cas9 protein gene. As the result, the transgenic lines 23216004 and 23216005 were further validated (Fig. 1).
The sequences of the primers were as bellow:
  • CAS9-F: 5’-atggactataaggaccacgacgg-3’
  • CAS9-R: 5’-ttgtcgcctcccagctgagacag-3’
Fig. 1 Detection results of PCR for gene Cas9.