IGEM:IMPERIAL/2007/Notebook/2007-9-14

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==Restriction Digest of Biobricks==
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#4 DNA samples were digested by SpeI and PstI
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#* Samples 5, 7, 24
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#1 DNA sample wasdigested by XbaI and PstI
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#* Samples 23
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#4 DNA samples were digested by EcoRI
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#* Samples 13, 25, 27b-e, T7, Kirsten's parts
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#4 μl of DNA was digested by 1 μl of enzymes at 37&deg;C for 1 hour<br>Exception: Sample 23 had 6 μl DNA and 2 μl enzymes
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Protocols can be found at [[IGEM:IMPERIAL/2007/Notebook/General_Protocols#Restriction_Digest|Restriction Digest]] in the general protocols page
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==Agarose Gel Electrophoresis of Biobricks==
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#Checked 9 Biobricks and 13 Digests on 1% agarose gel
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[[Image:ICGEMS Gel 14 9.png]]
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Conclusions:
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*Part 27 is confirmed to be faulty
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*T7 maxi-prep was not done properly
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*Kirsten's parts were most likely not there...
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Protocols can be found at [[IGEM:IMPERIAL/2007/Notebook/General_Protocols#Electrophoresis|Electrophoresis]] in the general protocols page
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==Gel Purification of Biobricks==
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#Gel was melted and run through column
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#Purified DNA was eluted with ddH<sub>2</sub>O
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*5. BBa_R0040 [ptet] (~2100 bp fragment)
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*7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment)
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*23. BBa_I13504 [GFP] (~800 bp fragment)
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*24. BBa_R0062 [pLux] (~2100 bp fragment)
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Protocols can be found at [[IGEM:IMPERIAL/2007/Notebook/General_Protocols#Gel_Purification|Gel Purification]] in the general protocols page

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Restriction Digest of Biobricks

  1. 4 DNA samples were digested by SpeI and PstI
    • Samples 5, 7, 24
  2. 1 DNA sample wasdigested by XbaI and PstI
    • Samples 23
  3. 4 DNA samples were digested by EcoRI
    • Samples 13, 25, 27b-e, T7, Kirsten's parts
  4. 4 μl of DNA was digested by 1 μl of enzymes at 37°C for 1 hour
    Exception: Sample 23 had 6 μl DNA and 2 μl enzymes

Protocols can be found at Restriction Digest in the general protocols page

Agarose Gel Electrophoresis of Biobricks

  1. Checked 9 Biobricks and 13 Digests on 1% agarose gel

Image:ICGEMS Gel 14 9.png

Conclusions:

  • Part 27 is confirmed to be faulty
  • T7 maxi-prep was not done properly
  • Kirsten's parts were most likely not there...

Protocols can be found at Electrophoresis in the general protocols page

Gel Purification of Biobricks

  1. Gel was melted and run through column
  2. Purified DNA was eluted with ddH2O

  • 5. BBa_R0040 [ptet] (~2100 bp fragment)
  • 7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment)
  • 23. BBa_I13504 [GFP] (~800 bp fragment)
  • 24. BBa_R0062 [pLux] (~2100 bp fragment)

Protocols can be found at Gel Purification in the general protocols page

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