User:Lu Wang/Notebook/Team Allergy/2010/07/28

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(Autocreate 2010/07/28 Entry for User:Lu_Wang/Notebook/Team_Allergy)
Current revision (15:00, 30 July 2010) (view source)
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==Entry title==
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==Protocol==
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Yesterday, we plated E. coli containing complete hpRNA for Ger, Bet v2, and LTP. Bet v1 did not ligate properly in the earlier Sense+Intron ligation.
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Today, we will extract the plasmids from the E. coli and prepare them for sequencing tomorrow. We will also do ligations with the false negatives (Sense + PDK ligate with Antisense).
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#Culture colonies
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#Miniprep
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#Nanodrop
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#Digest false negatives
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#Ligate
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#Transform
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==Results==
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'''Nanodrop'''
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The format is: Miniprep # , Name of Allergen, concentration in ng/μL of colony 1, concentration in ng/μL of colony 2.
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All these working ligations are with the PAL intron
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#9, LTP, 97.9, 94.1
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#11, LTP, 77.9, 94.8
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#25, Bet v2, 67.8, 97.8 (there was a mixup with colony 1 and LTP+PDK, but it is now fixed in our electronic notebooks)
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#28, Bet v2, 73.2, 83.6
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#36, Ger3, 76.1, 83.4
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#37, Ger3, 70.0, 75.3
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#38, Ger3, 90.4, 108.3
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*Transformed more sense+pdk parts for our false negatives (16,30,31)
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Protocol

Yesterday, we plated E. coli containing complete hpRNA for Ger, Bet v2, and LTP. Bet v1 did not ligate properly in the earlier Sense+Intron ligation.

Today, we will extract the plasmids from the E. coli and prepare them for sequencing tomorrow. We will also do ligations with the false negatives (Sense + PDK ligate with Antisense).

  1. Culture colonies
  2. Miniprep
  3. Nanodrop
  4. Digest false negatives
  5. Ligate
  6. Transform

Results

Nanodrop

The format is: Miniprep # , Name of Allergen, concentration in ng/μL of colony 1, concentration in ng/μL of colony 2. All these working ligations are with the PAL intron

  1. 9, LTP, 97.9, 94.1
  2. 11, LTP, 77.9, 94.8
  3. 25, Bet v2, 67.8, 97.8 (there was a mixup with colony 1 and LTP+PDK, but it is now fixed in our electronic notebooks)
  4. 28, Bet v2, 73.2, 83.6
  5. 36, Ger3, 76.1, 83.4
  6. 37, Ger3, 70.0, 75.3
  7. 38, Ger3, 90.4, 108.3
  • Transformed more sense+pdk parts for our false negatives (16,30,31)


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