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− | <a href=" | + | <a href="https://static.igem.org/mediawiki/2018/6/64/T--Munich--Phactory_Purifier.pdf" target="_blank">Filtering of Phage solution with Phactory purifier |
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− | <a href=" | + | <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> | ||
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− | Phage Encapsulation- | + | Phage Encapsulation-Stain of Capsules with DAPI |
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− | Phage Encapsulation-Sybr Gold | + | Phage Encapsulation-Sybr Gold Stain |
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− | Phage Encapsulation-Test of | + | Phage Encapsulation-Test of Cleaning Phages |
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− | <a href="https://static.igem.org/mediawiki/2018/2/23/T--Munich--AgarOverlayPlaqueAssay_.pdf" target="_blank">Plaque Assay</a> | + | <a href="https://static.igem.org/mediawiki/2018/2/23/T--Munich--AgarOverlayPlaqueAssay_.pdf" target="_blank">Plaque Assay</a> |
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− | <a href=" | + | <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> | ||
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− | Phage Encapsulation with Sybr Gold | + | Phage Encapsulation with Sybr Gold Staining |
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− | Test of | + | Test of Encapsulated Phages in HCl(aq) |
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− | Test of Phages | + | Test of Phages Droplets in SGF and SIF: Phage Encapsulation |
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− | <a href=" | + | <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> |
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Latest revision as of 01:20, 18 October 2018
Phage Encapsulation
2018/08/30Participants: | Johann Brenner, Keno Eilers |
Protocol: | Alginate Encapsulation | Notes: | No notes. | Results: | First Test of Encapsulation, Droplets: r=30 µM, polydisperse. |
Phage Encapsulation
30/08/2018Participants: | 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
2018/09/02Participants: | 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
2018/09/04Participants: | 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
2018/09/04Participants: | 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
2018/09/05Participants: | Johann Brenner, Keno Eilers |
Protocol: | Alginate Encapsulation | Notes: | No notes. | Results: | Test of transferring Droplets in Phage Buffer --> white sediment; CaPO4 crystals. |
Phage Encapsulation
2018/09/12Participants: | 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
2018/09/13Participants: | Johann Brenner |
Protocol: | Alginate Encapsulation | Notes: | No notes. | Results: | Design and print of new apparatus which holds dispenser needle in place. |
Phactory Purifier
2018/09/13Participants: | 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 - 2018/09/20Participants: | Johann Brenner, Keno Eilers |
Protocol: | Alginate Encapsulation | Notes: | No notes. | Results: | Test of different flow rates, enhanced reproducibility. |
Phage Encapsulation
2018/09/21 - 2018/10/01Participants: | 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
2018/10/02Participants: | 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
2018/10/04Participants: | 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
2018/10/04Participants: | 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
2018/10/06Participants: | 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)
2018/10/06Participants: | 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 - 2018/10/14Participants: | 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. |