Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/11 August 2015"

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='''Ligation of psb1c3+T7 promoter and Silk+GFP'''=
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{| class="wikitable" style="text-align:center; width:400px; height:200px;"
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|+
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|-
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! Component
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! Volume (out of 20uL)
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|-
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! T4 Ligase Reaction Buffer
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| 2 ul
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|-
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! Backbone (psb1c3+t7 promoter)
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| 0.8uL (50 ng)
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|-
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! Insert (silk + spyC)
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| 1.5 ul (146 ng)
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|-
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! H20
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| 14.7 ul
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|-
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! Ligase
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| 1uL
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|}
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*Program = 27C for 1 hour, heat inactivate for 10 minutes at 65C.
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=8/11=
 
=8/11=
 
===Protein Yield===
 
===Protein Yield===

Revision as of 22:02, 24 August 2015

Ligation of psb1c3+T7 promoter and Silk+GFP

Component Volume (out of 20uL)
T4 Ligase Reaction Buffer 2 ul
Backbone (psb1c3+t7 promoter) 0.8uL (50 ng)
Insert (silk + spyC) 1.5 ul (146 ng)
H20 14.7 ul
Ligase 1uL
  • Program = 27C for 1 hour, heat inactivate for 10 minutes at 65C.

8/11

Protein Yield

  • Yield from 8/7 silk purification
  • BCA assay
    • Based on the interpolated Absorbencies from the BCA assay the concentration of our 8/7 silk is approximately 0.78 mg/ml in around 12 ml of solution, giving a conservative 9 total mg of protein for a 300 ml culture.
  • This is an improvement over the 7/29 purification where we got around 1.6 mg/ml in only 0.75 ml of solution from a 100 ml culture.