Difference between revisions of "Team:Vilnius-Lithuania/Labjournal"
Line 126: | Line 126: | ||
<div class="row"> | <div class="row"> | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
− | Part: <strong> | + | Part: <strong>P15.1</strong> (BBa_) |
</div> | </div> | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
Line 141: | Line 141: | ||
<div class="col-md-12"> | <div class="col-md-12"> | ||
<h3><strong>About</strong></h3> | <h3><strong>About</strong></h3> | ||
− | <p> | + | <p>Cas3 with mutated restriction sites<br />L7+L15. Concentration - 261 ng/μL</p> |
</div> | </div> | ||
</div> | </div> | ||
Line 158: | Line 158: | ||
<tr> | <tr> | ||
<td>06 30</td> | <td>06 30</td> | ||
− | <td> | + | <td>P13<br />P7</td> |
− | <td> | + | <td>P13 and P7 transformation into DH5α.</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>07 01</td> | <td>07 01</td> | ||
<td></td> | <td></td> | ||
− | <td> | + | <td>Transformants are inoculated (4 minipreps each).</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
− | <td>07 | + | <td>07 02</td> |
− | <td> | + | <td></td> |
− | <td> | + | <td>Plasmid extraction. Concentrations:<br /> |
− | + | P7 – 174,1 ng/μL and 166 ng/μL.<br /> | |
− | + | P13 - 216 ng/μL and 131, 5 ng/μL. | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</td> | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>07 13</td> | ||
+ | <td>P13</td> | ||
+ | <td>Cas3 mutagenesis (with ST1/2, ST3/4, ST5/6 primers).</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>07 16</td> | ||
+ | <td>Mutated P13</td> | ||
+ | <td>Cas3 plasmids extraction.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>07 21-23</td> | ||
+ | <td>Mutated P13</td> | ||
+ | <td>Restriction with EcoRI, XbaI and PstI.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>07 27</td> | ||
+ | <td>Mutated P13</td> | ||
+ | <td>Cas3 mutagenesis.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>07 29</td> | ||
+ | <td>P7</td> | ||
+ | <td>Restriction with EcoRI and SpeI. L7 is produced.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>07 31</td> | ||
+ | <td>P13</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>08 02</td> | ||
+ | <td>Mutated P13</td> | ||
+ | <td>Mutated Cas3 restriction with EcoRI, XbaI and PstI.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>08 05</td> | ||
+ | <td>Mutated P13</td> | ||
+ | <td>Cas3 mutagenesis.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>08 07</td> | ||
+ | <td>Mutated P13</td> | ||
+ | <td>Mutated Cas3 restriction with PstI. Mutagenesis was successful. <br /> | ||
+ | L15 is produced.</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>08 11</td> | ||
+ | <td>L7<br /> L15</td> | ||
+ | <td>L7 and L15 ligation with pSB1C3 (EcoRI+XBal+FastAp).<br /> | ||
+ | P15.1 is produced.</td> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>P15.1</td> | ||
+ | <td>Sequencing.</td> | ||
</tr> | </tr> | ||
</tbody> | </tbody> | ||
Line 186: | Line 238: | ||
<div class="col-md-12"> | <div class="col-md-12"> | ||
<h3><strong>Conclusions and Results:</strong></h3> | <h3><strong>Conclusions and Results:</strong></h3> | ||
− | <p style="height: auto; margin: auto;">07 | + | <p style="height: auto; margin: auto;">07 21 – Mutagenesis was unsuccessful.<br/ > |
+ | 08 02 – successfully mutagenized EcoRI and XbaI sites.<br /> | ||
+ | 08 07 – successful mutagenesis confirmed (XbaI and EcoRI sites are mutated).<br /> | ||
+ | 08 07 – Successful mutagenesis (PstI, XbaI and EcoRI sites are mutated).</p> | ||
</div> | </div> | ||
</div> | </div> |
Revision as of 21:36, 7 September 2015
About
pLac+cI.
P10 and P11 plasmids are used. pLac (P11) and repressor cI (P10) ligation.
Date | Materials | Experiment |
---|---|---|
06 30 | P10 P11 |
P10 and P11 transformation into DH5α. |
07 01 | Bacteria inoculation (4 minipreps). | |
07 09 | P10 P11 |
Plasmids’ restriction (3 ug each). P10 with EcoRI and XBal (+FastAp). P11 with EcoRI and SpeI. Extraction from gel L10 and L11 are produced. Conc.: L11 – 2,4 ng/uL. |
Conclusions and Results:
07 14 – all of the colonies are successful, except third colony.
About
Cas3 with mutated restriction sites
L7+L15. Concentration - 261 ng/μL
Date | Materials | Experiment |
---|---|---|
06 30 | P13 P7 |
P13 and P7 transformation into DH5α. |
07 01 | Transformants are inoculated (4 minipreps each). | |
07 02 | Plasmid extraction. Concentrations: P7 – 174,1 ng/μL and 166 ng/μL. P13 - 216 ng/μL and 131, 5 ng/μL. |
|
07 13 | P13 | Cas3 mutagenesis (with ST1/2, ST3/4, ST5/6 primers). |
07 16 | Mutated P13 | Cas3 plasmids extraction. |
07 21-23 | Mutated P13 | Restriction with EcoRI, XbaI and PstI. |
07 27 | Mutated P13 | Cas3 mutagenesis. |
07 29 | P7 | Restriction with EcoRI and SpeI. L7 is produced. |
07 31 | P13 | |
08 02 | Mutated P13 | Mutated Cas3 restriction with EcoRI, XbaI and PstI. |
08 05 | Mutated P13 | Cas3 mutagenesis. |
08 07 | Mutated P13 | Mutated Cas3 restriction with PstI. Mutagenesis was successful. L15 is produced. |
08 11 | L7 L15 |
L7 and L15 ligation with pSB1C3 (EcoRI+XBal+FastAp). P15.1 is produced. |
P15.1 | Sequencing. |
Conclusions and Results:
07 21 – Mutagenesis was unsuccessful.
08 02 – successfully mutagenized EcoRI and XbaI sites.
08 07 – successful mutagenesis confirmed (XbaI and EcoRI sites are mutated).
08 07 – Successful mutagenesis (PstI, XbaI and EcoRI sites are mutated).
About
pLac+cI.
P10 and P11 plasmids are used. pLac (P11) and repressor cI (P10) ligation.
Date | Materials | Experiment |
---|---|---|
06 30 | P10 P11 |
P10 and P11 transformation into DH5α. |
07 01 | Bacteria inoculation (4 minipreps). | |
07 09 | P10 P11 |
Plasmids’ restriction (3 ug each). P10 with EcoRI and XBal (+FastAp). P11 with EcoRI and SpeI. Extraction from gel L10 and L11 are produced. Conc.: L11 – 2,4 ng/uL. |
Conclusions and Results:
07 14 – all of the colonies are successful, except third colony.
About
pLac+cI.
P10 and P11 plasmids are used. pLac (P11) and repressor cI (P10) ligation.
Date | Materials | Experiment |
---|---|---|
06 30 | P10 P11 |
P10 and P11 transformation into DH5α. |
07 01 | Bacteria inoculation (4 minipreps). | |
07 09 | P10 P11 |
Plasmids’ restriction (3 ug each). P10 with EcoRI and XBal (+FastAp). P11 with EcoRI and SpeI. Extraction from gel L10 and L11 are produced. Conc.: L11 – 2,4 ng/uL. |
Conclusions and Results:
07 14 – all of the colonies are successful, except third colony.
About
pLac+cI.
P10 and P11 plasmids are used. pLac (P11) and repressor cI (P10) ligation.
Date | Materials | Experiment |
---|---|---|
06 30 | P10 P11 |
P10 and P11 transformation into DH5α. |
07 01 | Bacteria inoculation (4 minipreps). | |
07 09 | P10 P11 |
Plasmids’ restriction (3 ug each). P10 with EcoRI and XBal (+FastAp). P11 with EcoRI and SpeI. Extraction from gel L10 and L11 are produced. Conc.: L11 – 2,4 ng/uL. |
Conclusions and Results:
07 14 – all of the colonies are successful, except third colony.