Team:UNIPV-Pavia/Notebook/Week12

From 2008.igem.org

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{|
{|
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|[[Image:pv_colonypcr_27_b.jpg|thumb|300px|left|Marker 1Kb, blank, BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-'''BBa_R0040''' (6 colonies), BBa_B0030-BBa_C0061-BBa_B1006-'''BBa_R0062'''-BBa_B0030-BBa_E0040-BBa_B1006 (7 colonies)]]
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|[[Image:pv_colonypcr_27_b.jpg|thumb|300px|left|Marker 1Kb, blank, BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-'''BBa_R0040''' (6 colonies), b (7 colonies)]]
|}
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*Gel result:
*Gel result:
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**BBa_B0030-BBa_C0061-BBa_B1006-'''BBa_R0062'''-BBa_B0030-BBa_E0040-BBa_B1006 (2nd colony)
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**b (2nd colony, but it was not pure. We decided to prepare single colonies plate for b)
**BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-'''BBa_R0010''' (1st colony)
**BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-'''BBa_R0010''' (1st colony)
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*Gel run/cut/gel extraction.
*Gel run/cut/gel extraction.
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 +
*We infected 9 ml of LB + Amp with 30 µl of BBa_C0062, 12, 22, 30, 27(2nd col), b(1st col)
*Single colonies plates for b, c, d.
*Single colonies plates for b, c, d.
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*We infected 9 ml of LB + Amp with 30
+
'''08/4/08'''
 +
<br>
 +
*Glycerol stocks/miniprep for BBa_C0062, 12, 22, 30, 27(2), b(1)

Revision as of 10:53, 22 September 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 11: 08/4/08 - 08/8/08

08/4/08

  • Plasmid digestion for:
BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006 (E-S) BBa_R0040 (E-X)
  • Gel run/cut/gel extraction.
  • Ligation: BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040. We incubated ligation at 16°C overnight.
  • We had 5 plates to screen with colony PCR:
    • BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E1010-BBa_B1006 (that we call "a")
    • BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E0040-BBa_B1006 (that we call "b")
    • BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079-BBa_B0030-BBa_E0040-BBa_B1006 (that we call "c")
    • BBa_B0030-BBa_C0061-BBa_B0030-BBa_C0079-BBa_B1006-BBa_R0079-BBa_B0030-BBa_E0040-BBa_B1006 (that we call "d")
    • BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040
  • Last week BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040 colony PCR gave a bad result. For this reason, we decided to perform colony PCR only for:
    • BBa_B0030-BBa_C0061-BBa_B1006-BBa_R0062-BBa_B0030-BBa_E0040-BBa_B1006 (7 colonies)
    • BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-BBa_R0010 (6 colonies)
Marker 1Kb, blank, BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040 (6 colonies), b (7 colonies)
  • Gel result:
    • b (2nd colony, but it was not pure. We decided to prepare single colonies plate for b)
    • BBa_J23100-BBa_B0030-BBa_C0012-BBa_B1006-BBa_R0010 (1st colony)

08/4/08

  • Plasmid digestion for:
BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006 (E-S) BBa_R0040 (E-X)
  • Gel run/cut/gel extraction.
  • We infected 9 ml of LB + Amp with 30 µl of BBa_C0062, 12, 22, 30, 27(2nd col), b(1st col)
  • Single colonies plates for b, c, d.

08/4/08

  • Glycerol stocks/miniprep for BBa_C0062, 12, 22, 30, 27(2), b(1)