Difference between revisions of "Team:UCLA/Notebook/Materials"

 
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</li><li>3rd change: 7:00 pm, May 13
 
</li><li>3rd change: 7:00 pm, May 13
 
</li><li>4th change: 2:00 pm, May 14
 
</li><li>4th change: 2:00 pm, May 14
 +
</li><li>5th change: 12:00 pm, May 15
 
<ul><li>Note: no leakage observed. Good sign so far.
 
<ul><li>Note: no leakage observed. Good sign so far.
 
</li></ul>
 
</li></ul>
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<ul><li>note to self: just use a more powerful centrifuge. It isn't worth it to split up the sample into microtubes to spin in our table top centrifuge.
 
<ul><li>note to self: just use a more powerful centrifuge. It isn't worth it to split up the sample into microtubes to spin in our table top centrifuge.
 
</li><li>ended up with 25 mL of silk
 
</li><li>ended up with 25 mL of silk
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</li><li>pipetted out 500 uL to dry to see our silk concentration
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</li><li>pipetted out 100 uL to dry to see our silk concentration
 
<ul><li>I am expecting a very low concentration, due to the way that our dialysis was leaking.
 
<ul><li>I am expecting a very low concentration, due to the way that our dialysis was leaking.
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</li><li>the solution ended up having 0.006 grams of silk.
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</li><li>This would be a 6% w/v solution. Surprisingly high, given the fact that it was leaking.
 
</li></ul>
 
</li></ul>
 
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<details><summary>May 13</summary>
 
<details><summary>May 13</summary>
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<ul><li>20% w/v dope extruded into 70% ethanol succeeded
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<ul><li>nominal extrusion rate of 16 uL/min
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</li><li>I say nominal because the screw drive clearly isn’t advancing
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<ul><li>I see now why professional fiber spinners use piston pressure instead of a drive screw syringe pump
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</li></ul>
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</li><li>I had to pull the coagulating mass out of the way of the dope that was continuing to advance out of the PEEK tubing
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<ul><li>hence, at some points the fiber is thinner (but still intact)
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</li><li>nevertheless, even without pulling on the dope, a fiber would still form right out of the tubing
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</li></ul>
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</li><li>Some confounding variables:
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<ul><li>I have no idea what pressure the pump is applying to the syringe since the drive block is not moving at all
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</li><li>Time may be a variable: this had been pumping for a long time before this particular experiment. Maybe all of the air bubbles from yesterday have already been purged?
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</li></ul>
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</li><li>Things to take care of next:
 +
<ul><li>construct a robust set up where dope coming out of the tube will have adequate space so that it doesn’t hinder the dope coming out of the peek tubing
 +
</li><li>try it on silk!
 +
</li><li>construct a post-spin stretch system. This will alleviate the problem of coagulated dope interfering with dope that’s still coming out
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<ul><li>additionally, we’ll need to have this set up anyway for silk
 +
</li></ul>
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</li></ul>
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</li></ul>
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</li><li>Once again, tried 20% w/v dope extruded into 70% ethanol, this time with the syringe pump oriented vertically
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<ul><li>gravity pulled already forming fibers out of the way, allowing fibers to continue to form.
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</li></ul>
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</li></ul>
  
 
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<details><summary>Day 3</summary>
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<details><summary>May 14</summary>
 
<ul>
 
<ul>
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<li>PCR</li>
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<li>Degummed more silk (2.5 grams)</li>
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<li>Ran gel</li>
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<li>pipetted 100 uL of the dialyzed silk from Tuesday to determine concentration</li>
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<li>Restriction digest</li>
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<li>Spun more cellulose acetate (same conditions from yesterday)</li>
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<li>Had a slice of pizza</li>
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<li>prepared LiBr solution (16 mL, 9.3M)</li>
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</ul>
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<li>took light microscope images of the fibers obtained yesterday</li>
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<img src="https://static.igem.org/mediawiki/2015/d/df/UCLA_14May2015_Cellulose_acetate_20pct_20x_2.jpg" alt = "cellulose acetate fiber 20% at 20x mag" height="500" >
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<ul><li>The spun fiber is approximately the same diameter as the PEEK tubing. For reference, the inner diameter of the PEEK tubing is 127 um.</li></ul>
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<img src="https://static.igem.org/mediawiki/2015/b/b0/UCLA_14May2015_Cell_acetate_20pct_20x_2.1.jpg" alt ="cellulose acetate fiber 20% at 20x mag" height = "500">
 +
<ul><li>However, note that the fiber is non-uniform. This is an image of a different spot along the same fiber. This part of the fiber is likely thicker because the dope bent while coagulating in the bath.</li></ul>
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== Goals ==
 +
*Wet-Spin fibers from mixtures of silkworm silk and recombinant functionalized spider silk
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*Wet-Spin fibers from recombinant honeybee silk
 +
*Construct a post-spin stretch apparatus
 +
*Characterize all spun fibers
 +
**Elastic modulus
 +
**Yield stress
 +
**Beta-sheet content (FTIR)
  
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==Entries for every day of work are available through the calendars below==
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*Create films from recombinant silk
 +
*Characterize recombinant silk films
 +
**Elastic modulus
 +
**Yield Stress
 +
 
 +
==Achievements==
 +
*May 13: completed our practice spins on cellulose acetate!
 +
**[https://2015.igem.org/Team:UCLA/Notebook/Materials/14_May_2015 Check out microscopy on our first fibers]
 +
 
 +
*May 28: we reconstituted silk solution into fibers!
 +
**[https://2015.igem.org/Team:UCLA/Notebook/Materials/29_May_2015 Preliminary images of our first reconstituted silk fibers]
 +
 
 +
*July 9: preliminary success with co-spinning b.mori silk w/ recombinant Tamura silk-GFP hybrid
 +
**[https://2015.igem.org/Team:UCLA/Notebook/Materials/9_July_2015 Some microscopy images here]
 +
 
 +
*July 14: a long contiguous co-spin:
 +
**[https://2015.igem.org/Team:UCLA/Notebook/Materials/14_July_2015 100 uL of dope yields a fairly long thread!]
 +
 
 +
*August 19: co-spin with the automated stretcher:
 +
**[https://2015.igem.org/Team:UCLA/Notebook/Materials/18_August_2015]
 +
 
 +
 
 +
== What to accomplish next ==
 +
*Spin fibers from the recombinant spider silk dopes
 +
 
 +
==Protocols==
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*[Team:UCLA/Notebook/Materials/degumming | Silk Degumming]
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*[Team:UCLA/Notebook/Materials/solubilization | Silk Solubilization]
 +
*[https://2015.igem.org/Team:UCLA/Notebook/Materials/Dialysis_LiBr Dialysis to remove LiBr]
 +
*https://2015.igem.org/Team:UCLA/Notebook/Materials/Dialysis_Concentrate Dialysis to concentrate silk solution]
 +
*[https://2015.igem.org/Team:UCLA/Notebook/Materials/WetSpinning Wet Spinning]
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*[https://2015.igem.org/Team:UCLA/Notebook/Materials/Instron Using the Instron]
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*[https://2015.igem.org/Team:UCLA/Notebook/Materials/Lyophilizer Using the lyophilizer]
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*[https://2015.igem.org/Team:UCLA/Notebook/Materials/HFIP Dissolving silk in HFIP]
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*[https://2015.igem.org/Team:UCLA/Notebook/Materials/Quick_Reference Quick Reference]
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== Raw notebook entries ==
 
{{#calendar: year=2015 | month= may | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
 
{{#calendar: year=2015 | month= may | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
 
{{#calendar: year=2015 | month= jun | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
 
{{#calendar: year=2015 | month= jun | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
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{{#calendar: year=2015 | month= aug | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
 
{{#calendar: year=2015 | month= aug | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
 
{{#calendar: year=2015 | month= sep | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
 
{{#calendar: year=2015 | month= sep | title = Team:UCLA/Notebook/Materials | query=preload=Template:UCLA}}
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[[File:UCLAiGEM_2015_Silk_Processing_flowchart.svg|1000 px]]

Latest revision as of 03:01, 19 September 2015

iGEM UCLA




Materials Processing Notebook



Goals

  • Wet-Spin fibers from mixtures of silkworm silk and recombinant functionalized spider silk
  • Wet-Spin fibers from recombinant honeybee silk
  • Construct a post-spin stretch apparatus
  • Characterize all spun fibers
    • Elastic modulus
    • Yield stress
    • Beta-sheet content (FTIR)
  • Create films from recombinant silk
  • Characterize recombinant silk films
    • Elastic modulus
    • Yield Stress

Achievements

  • August 19: co-spin with the automated stretcher:


What to accomplish next

  • Spin fibers from the recombinant spider silk dopes

Protocols

Raw notebook entries

May
M T W T F S S
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31
June
M T W T F S S
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30
July
M T W T F S S
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
M T W T F S S
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
M T W T F S S
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
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