Difference between revisions of "Team:Munich/capsulation.html"

(Created page with "<html> <div class="event-info"> <h4> Phage Encapsulation </h4> <em> 2018/08/30 </em> <table class=”table table-borderless”> <tr> <td>Participants:</td> <td>Johann Brenner,...")
 
 
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<h4>
 
<h4>
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Phactory purificator
+
Phactory Purifier
 
</h4>
 
</h4>
 
<em>
 
<em>
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<td>Protocol:</td>
 
<td>Protocol:</td>
 
<td>
 
<td>
−
<a href="HERE" target="_blank">NAME</a>,
+
<a href="https://static.igem.org/mediawiki/2018/6/64/T--Munich--Phactory_Purifier.pdf" target="_blank">Filtering of Phage solution with Phactory purifier
 +
</a>
 
</td>
 
</td>
 
</tr>
 
</tr>
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<h4>
 
<h4>
−
Phactory purificator
+
Phactory Purifier
 
</h4>
 
</h4>
 
<em>
 
<em>
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<td>Protocol:</td>
 
<td>Protocol:</td>
 
<td>
 
<td>
−
<a href="HERE" target="_blank">NAME</a>,
+
<a href="https://static.igem.org/mediawiki/2018/6/64/T--Munich--Phactory_Purifier.pdf" target="_blank">Filtering of Phage solution with Phactory purifier
 +
</a>
 
</td>
 
</td>
 
</tr>
 
</tr>
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<h4>
 
<h4>
−
Phage Encapsulation-stain of capsules with DAPI
+
Phage Encapsulation-Stain of Capsules with DAPI
 
</h4>
 
</h4>
 
<em>
 
<em>
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<td>
 
<td>
 
Test of different flow rates, enhanced reproducibility.  
 
Test of different flow rates, enhanced reproducibility.  
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Phage Encapsulation
 +
</h4>
 +
<em>
 +
2018/09/21 - 2018/10/01
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
<a href="https://static.igem.org/mediawiki/2018/8/85/T--Munich--AlginatEncapsulation.pdf" target="_blank">Alginate Encapsulation</a>
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Test of different Alginate/low viscosity Alginate ratio 1.8% Alginate + 0.2%low viscosity gave the best results.
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Phage Encapsulation-Sybr Gold Stain
 +
</h4>
 +
<em>
 +
2018/10/02
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
<a href="https://static.igem.org/mediawiki/2018/8/85/T--Munich--AlginatEncapsulation.pdf" target="_blank">Alginate Encapsulation</a>
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Sybr Gold stained Phages are clearly visible, next time wash away of un bound Sybr Gold.
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Phage Encapsulation-Test of Cleaning Phages
 +
</h4>
 +
<em>
 +
2018/10/04
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
<a href="https://static.igem.org/mediawiki/2018/2/23/T--Munich--AgarOverlayPlaqueAssay_.pdf" target="_blank">Plaque Assay</a>
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Phages (in PBS) were centrifuged with 30 K centrifugal filters and stored in 10mM MgSO4 concentration of phages can be increased.
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Phactory Purifier
 +
</h4>
 +
<em>
 +
2018/10/04
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
<a href="https://static.igem.org/mediawiki/2018/6/64/T--Munich--Phactory_Purifier.pdf" target="_blank">Filtering of Phage solution with Phactory purifier
 +
</a>
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Spot test positive for several fractions,  positive fractions were concentrated with centrifugal filters.
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Phage Encapsulation with Sybr Gold Staining
 +
</h4>
 +
<em>
 +
2018/10/06
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
<a href="https://static.igem.org/mediawiki/2018/8/85/T--Munich--AlginatEncapsulation.pdf" target="_blank">Alginate Encapsulation</a>
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Clearly visible fluorescent signal with less background than before.
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Test of Encapsulated Phages in HCl(aq)
 +
</h4>
 +
<em>
 +
2018/10/06
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
No protocol.
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Phages were monitored over 7h in HCl ph1, microspheres stay the same size over the time.
 +
</td>
 +
</tr>
 +
</table>
 +
 +
<h4>
 +
Test of Phages Droplets in SGF and SIF: Phage Encapsulation
 +
</h4>
 +
<em>
 +
2018/10/07 - 2018/10/14
 +
</em>
 +
<table class=”table table-borderless”>
 +
<tr>
 +
<td>Participants:</td>
 +
<td>Johann Brenner, Keno Eilers</td>
 +
</tr>
 +
<tr>
 +
<td>Protocol:</td>
 +
<td>
 +
<a href="https://static.igem.org/mediawiki/2018/9/9a/T--Munich--Results_Protocl_EncapsulatedPhagesinSGFandSIF.pdf" target="_blank">Validation of encapsulated T4 phage in simulated gastric fluid (SGF) and Simulated intestinal fluid (SIF) </a>
 +
</td>
 +
</tr>
 +
<td>Notes:</td>
 +
        <td>
 +
No notes.
 +
      </td>
 +
</tr>
 +
<td>Results:</td>
 +
<td>
 +
Capsules show enhanced stability in SGF and high release of active phages in SIF.
 
</td>
 
</td>
 
</tr>
 
</tr>

Latest revision as of 01:20, 18 October 2018

Phage Encapsulation

2018/08/30
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: First Test of Encapsulation, Droplets: r=30 µM, polydisperse.

Phage Encapsulation

30/08/2018
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Results: Design of own apparatus and first 3D testprints, filament evaluation (PLA, PMMA).

Phage Encapsulation

2018/09/02
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: First test of 3D print, needs some updates for better fitting, PMMA seems to be best material.

Phage Encapsulation

2018/09/04
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Works better now, better assembly, Droplets look good, but a lot of aggregate, next time more CaCl2 solution.

Phactory Purifier

2018/09/04
Participants: Johann Brenner, Keno Eilers
Protocol: Filtering of Phage solution with Phactory purifier
Notes: No notes.
Results: Spot test positive for fractions 13-23.

Phage Encapsulation

2018/09/05
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Test of transferring Droplets in Phage Buffer --> white sediment; CaPO4 crystals.

Phage Encapsulation

2018/09/12
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Alginate solution with10 mM MgSO4, different batches with different flow rates and N2 pressure, lack of reproducibility.

Phage Encapsulation

2018/09/13
Participants: Johann Brenner
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Design and print of new apparatus which holds dispenser needle in place.

Phactory Purifier

2018/09/13
Participants: Johann Brenner, Keno Eilers
Protocol: Filtering of Phage solution with Phactory purifier
Notes: No notes.
Results: Spot test positive for fractions 10-23.

Phage Encapsulation-Stain of Capsules with DAPI

2018/09/14 - 2018/09/20
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Test of different flow rates, enhanced reproducibility.

Phage Encapsulation

2018/09/21 - 2018/10/01
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Test of different Alginate/low viscosity Alginate ratio 1.8% Alginate + 0.2%low viscosity gave the best results.

Phage Encapsulation-Sybr Gold Stain

2018/10/02
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Sybr Gold stained Phages are clearly visible, next time wash away of un bound Sybr Gold.

Phage Encapsulation-Test of Cleaning Phages

2018/10/04
Participants: Johann Brenner, Keno Eilers
Protocol: Plaque Assay
Notes: No notes.
Results: Phages (in PBS) were centrifuged with 30 K centrifugal filters and stored in 10mM MgSO4 concentration of phages can be increased.

Phactory Purifier

2018/10/04
Participants: Johann Brenner, Keno Eilers
Protocol: Filtering of Phage solution with Phactory purifier
Notes: No notes.
Results: Spot test positive for several fractions, positive fractions were concentrated with centrifugal filters.

Phage Encapsulation with Sybr Gold Staining

2018/10/06
Participants: Johann Brenner, Keno Eilers
Protocol: Alginate Encapsulation
Notes: No notes.
Results: Clearly visible fluorescent signal with less background than before.

Test of Encapsulated Phages in HCl(aq)

2018/10/06
Participants: Johann Brenner, Keno Eilers
Protocol: No protocol.
Notes: No notes.
Results: Phages were monitored over 7h in HCl ph1, microspheres stay the same size over the time.

Test of Phages Droplets in SGF and SIF: Phage Encapsulation

2018/10/07 - 2018/10/14
Participants: Johann Brenner, Keno Eilers
Protocol: Validation of encapsulated T4 phage in simulated gastric fluid (SGF) and Simulated intestinal fluid (SIF)
Notes: No notes.
Results: Capsules show enhanced stability in SGF and high release of active phages in SIF.