Team:UNIPV-Pavia/Notebook/Week7

From 2008.igem.org

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'''07/1/08'''
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'''07/2/08'''
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*We transformed ligations (5 µl) and plated transformed bacteria.
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 +
*We also transformed BBa_B1006(E-X), BBa_J23100-'''BBa_B0030'''(S-P) and '''BBa_R0051'''-BBa_B0030 (1 µl) and plated transformed bacteria to estimate background noise.
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*We received sequencing results for:
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**'''BBa_B0030'''-BBa_E0040
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**'''BBa_B0030'''-BBa_C0051
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**'''BBa_B0030'''-BBa_E1010
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*All the sequences were correct!
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*We infected 9 ml LB + Amp with 30 µl of:
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{|cellpadding="20px"
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|BBa_B0030
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|'''BBa_B0030'''-BBa_E0040
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|'''BBa_B0030'''-BBa_C0051
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|-
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|BBa_B1006
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|'''BBa_B0030'''-BBa_E1010
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|}
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*glycerol stocks.

Revision as of 22:04, 13 July 2008


Home.jpg Home Unipv logo.jpg The Team And.jpg The Project Safety.jpg Biological Safety Dna.png Parts Submitted to the Registry
Laptop.jpg Dry Lab Pipette.jpg Wet Lab Math.gif Modeling Note.jpg Protocols Notebook.gif Activity Notebook



Notebook



Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7
Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14



Week 7: 06/30/08 - 06/5/08

06/30/08

  • We received sequencing results for BBa_B0030-BBa_C0061 (4th colony and 7th colony): sequences were correct! We decided to keep the 4th colony.
  • Colony PCR for BBa_J23100-BBa_E0240 and BBa_B0030-BBa_C0061: 10 colonies for every plate. Gel showed many working colonies: we chose first colonies for the two ligations.
Colony PCR for BBa_J23100-BBa_E0240, BBa_B0030-BBa_E0061: Marker and 10 colonies for each ligation
  • We infected 9 ml LB + Amp with 30 µl of:
BBa_C0012 BBa_B1006 BBa_R0051-BBa_B0030 BBa_B0030-BBa_C0061
BBa_C0062 BBa_J23100-BBa_E0240 (1) BBa_B0030-BBa_C0078 (1) BBa_J23100-BBa_B0030
BBa_C0040 BBa_I15010
  • Tomorrow we will be ready to perform 5 ligations!



07/1/08

  • Glycerol stocks for:
BBa_C0012 BBa_B1006 BBa_R0051-BBa_B0030 BBa_B0030-BBa_C0061
BBa_C0062 BBa_J23100-BBa_E0240 (1) BBa_B0030-BBa_C0078 (1) BBa_J23100-BBa_B0030
BBa_C0040 BBa_I15010
  • Miniprep for all these parts.
  • We sent BBa_J23100-BBa_E0240 (1) and BBa_B0030-BBa_C0078 (1) to Primm for sequencing.
  • We performed digestion for:
BBa_C0012 (X-P) BBa_B1006 (E-X) BBa_R0051-BBa_B0030 (S-P) BBa_B0030-BBa_C0061 (E-S)
BBa_C0062 (X-P) BBa_C0040 (X-P) BBa_I15010 (X-P) BBa_J23100-BBa_B0030 (S-P)
  • Gel run/cut for:
BBa_C0012 (X-P) BBa_C0062 (X-P) BBa_B0030-BBa_C0061 (E-S)
BBa_C0040 (X-P) BBa_I15010 (X-P)
  • Gel extraction for these 5 parts.
  • DNA precipitation with sodium acetate for:
BBa_B1006 (E-X) BBa_R0051-BBa_B0030 (S-P) BBa_J23100-BBa_B0030 (S-P)
  • Ligation:
    • BBa_J23100-BBa_B0030-BBa_C0012
    • BBa_J23100-BBa_B0030-BBa_C0040
    • BBa_J23100-BBa_B0030-BBa_I15010
    • BBa_R0051-BBa_B0030-BBa_C0062
    • BBa_B0030-BBa_C0061-BBa_B1006
  • We incubated ligation reaction at 16°C overnight.



07/2/08

  • We transformed ligations (5 µl) and plated transformed bacteria.
  • We also transformed BBa_B1006(E-X), BBa_J23100-BBa_B0030(S-P) and BBa_R0051-BBa_B0030 (1 µl) and plated transformed bacteria to estimate background noise.
  • We received sequencing results for:
    • BBa_B0030-BBa_E0040
    • BBa_B0030-BBa_C0051
    • BBa_B0030-BBa_E1010
  • All the sequences were correct!
  • We infected 9 ml LB + Amp with 30 µl of:
BBa_B0030 BBa_B0030-BBa_E0040 BBa_B0030-BBa_C0051
BBa_B1006 BBa_B0030-BBa_E1010
  • glycerol stocks.
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