Team:UNIPV-Pavia/Notebook/Week1

From 2008.igem.org

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*We plated transformed bacteria and incubated them at 37°C overnight.
*We plated transformed bacteria and incubated them at 37°C overnight.
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<br><br>
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'''05/21/08'''
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<br>
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*Only BBa_I15009 and BBa_C0078 plates showed colonies and for the remaining 6 plates there were no colonies again.
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*We picked up one colony from BBa_I15009 and BBa_C0078 plates to grow 5 ml cultures of transformed bacteria overnight.
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*We re-cut paper spots for BBa_R0051, BBa_I14032, BBa_I15008, BBa_I15010, BBa_C0161, BBa_C0179 and resuspended them again in 10 l of warmed TE buffer.
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*We repeated the transformation for these 6 parts using 4 µl of DNA in TE.
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*We plated transformed bacteria and incubated them at 37°C overnight.
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*We prepared 14 glycerol stocks taking 800 µl from 5 ml cultures containing:
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<table width="100%">
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<tr><td>
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{|cellpadding="20"
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|BBa_R0079
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|BBa_R0062
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|BBa_R0040
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|BBa_R0082
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|-
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|BBa_J23100
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|BBa_C0062
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|BBa_C0051
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|BBa_C0012
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|-
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|BBa_C0040
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|BBa_E0040
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|BBa_E1010
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|BBa_B0030
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|-
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|BBa_B1006
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|BBa_E0240
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|}
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</td>
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<td align="center">
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{|
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|[[Image:pv_red_colonies.jpg|thumb|300px|left|BBa_J23100 red culture and BBa_B0030 normal color colonies]]
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|}
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</td></tr>
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</table>

Revision as of 10:45, 28 May 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 1: 05/19/08 - May 05/23/08

05/19/08

  • Let’s start our IGEM 2008 experience! At first, we broke the punch tool…:)
  • We used a scalpel to cut and resuspend the following 22 paper spots:
BBa_I14032 BBa_R0079 BBa_R0062 BBa_R0040
BBa_R0082 BBa_R0051 BBa_J23100 BBa_C0161
BBa_C0062 BBa_C0078 BBa_C0179 BBa_C0051
BBa_C0012 BBa_C0040 BBa_I15010 BBa_I15008
BBa_I15009 BBa_E0040 BBa_E1010 BBa_B0030
BBa_B1006 BBa_E0240
Registry of Standard Parts 2008 in our lab
  • We used tweezers to put the cut paper into tubes containing 10 μl of warmed TE buffer.
  • We transformed 60 µl of TOP10 E. coli with 4 µl of DNA in TE for all 22 parts, plated transformed bacteria and incubated overnight at 37°C.
  • We used LB medium previously prepared, with the suitable antibiotic added.



05/20/08

  • After overnight incubation, the following 14 plates showed colonies:
BBa_R0079 BBa_R0062 BBa_R0040 BBa_R0082
BBa_J23100 BBa_C0062 BBa_C0051 BBa_C0012
BBa_C0040 BBa_E0040 BBa_E1010 BBa_B0030
BBa_B1006 BBa_E0240
BBa_E0240
  • While the following plates did not:
BBa_I14032 BBa_R0051 BBa_C0161 BBa_C0078
BBa_C0179 BBa_I15008 BBa_I15009 BBa_I15010


  • Plate containing BBa_J23100 showed red colonies, as we expected: this part is inserted into plasmid BBa_J61002 which places the RFP downstream of the inserted part, which is a constitutive promoter.
  • We picked up one colony from every working plate to grow 5 ml cultures of transformed bacteria overnight.
  • We re-transformed 60 µl of TOP10 with the remaining 6 µl of DNA in TE for BBa_I14032, BBa_R0051, BBa_I15008, BBa_I15009, BBa_I15010, BBa_C0161, BBa_C0078, BBa_C0179.
  • We plated transformed bacteria and incubated them at 37°C overnight.



05/21/08

  • Only BBa_I15009 and BBa_C0078 plates showed colonies and for the remaining 6 plates there were no colonies again.
  • We picked up one colony from BBa_I15009 and BBa_C0078 plates to grow 5 ml cultures of transformed bacteria overnight.
  • We re-cut paper spots for BBa_R0051, BBa_I14032, BBa_I15008, BBa_I15010, BBa_C0161, BBa_C0179 and resuspended them again in 10 l of warmed TE buffer.
  • We repeated the transformation for these 6 parts using 4 µl of DNA in TE.
  • We plated transformed bacteria and incubated them at 37°C overnight.
  • We prepared 14 glycerol stocks taking 800 µl from 5 ml cultures containing:
BBa_R0079 BBa_R0062 BBa_R0040 BBa_R0082
BBa_J23100 BBa_C0062 BBa_C0051 BBa_C0012
BBa_C0040 BBa_E0040 BBa_E1010 BBa_B0030
BBa_B1006 BBa_E0240
File:Pv red colonies.jpg
BBa_J23100 red culture and BBa_B0030 normal color colonies
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