Team:UNIPV-Pavia/Notebook/Week10

From 2008.igem.org

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'''07/21/08'''
'''07/21/08'''
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*We transformed 1 µl of the 6 ligations stored at -20°C. We plated transformed bacteria and incubated plates at 37°C overnight.
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*Colony PCR for BBa_B0030-BBa_C0078-BBa_B1006-'''BBa_R0079''': 6 colonies from single colonies plate.
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*Plasmid digestion for:
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{|
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{|cellpadding="20px"
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|[[Image:pv_colonypcr_25_single_col.jpg|thumb|300px|left|BBa_B0030-BBa_C0078-BBa_B1006-'''BBa_R0079''']]
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|BBa_R0079 (E-X)
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|BBa_B0030-BBa_C0078-'''BBa_B1006''' (E-S)
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|}
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*Gel run/cut/gel extraction for the 2 parts.
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*Gel results: all the 6 picked colonies were true positive! we chose the 5th colont to grow a 9 ml overnight culture.
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*Ligation: BBa_B0030-BBa_C0078-BBa_B1006-'''BBa_R0079'''.
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*We also infected 9 ml LB + Amp with 30 µl of BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-'''BBa_R0040''' and '''BBa_B0030'''-BBa_C0061 glycerol stocks to grow two overnight cultures.
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*We incubated ligation at 16°C overnight.
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*Ligation;
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'''07/15/08'''
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**
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**
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*We transformed 1 µl of BBa_B0030-BBa_C0078-BBa_B1006-'''BBa_R0079''' ligation and plated transformed bacteria. We incubated the plate at 37°C overnight.
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*All the 6 overnight plates showed colonies. We performed colony PCR for (we decided to pick up more colonies for the ligations with long inserts):
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#BBa_J23100-'''BBa_B0030''' (S-P) -BBa_C0012 (standard ligation) - 10 colonies
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#'''pSB1AK3'''-BBa_J23100-BBa_B0030-BBa_C0012 (double ligation) - 7 colonies
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#BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-'''BBa_R0040''' - 3 colonies
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#BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006-'''BBa_R0062''' - 3 colonies
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#BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-'''BBa_B1006''' - 10 colonies
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#BBa_B0030-BBa_I15009-'''BBa_B1006''' - 3 colonies
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{|
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|[[Image:pv_colonypcr_15_7_08.jpg|thumb|300px|left|Picked up colonies in LB + Amp]]
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Revision as of 13:08, 27 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 10: 07/21/08 - 07/25/08

07/21/08

  • Colony PCR for BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079: 6 colonies from single colonies plate.
BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079
  • Gel results: all the 6 picked colonies were true positive! we chose the 5th colont to grow a 9 ml overnight culture.
  • We also infected 9 ml LB + Amp with 30 µl of BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040 and BBa_B0030-BBa_C0061 glycerol stocks to grow two overnight cultures.
  • Ligation;
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