Team:UNIPV-Pavia/Notebook/Week1

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(Difference between revisions)
Line 71: Line 71:
*We used LB medium previously prepared, with the suitable antibiotic added.
*We used LB medium previously prepared, with the suitable antibiotic added.
 +
 +
'''05/20/08'''
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<br>
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*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
 +
|}
 +
 +
*While the following plates did not:
 +
 +
{|
 +
|BBa_I14032
 +
|BBa_R0051
 +
|BBa_C0161
 +
|-
 +
|BBa_C0078
 +
|BBa_C0179
 +
|BBa_I15008
 +
|-
 +
|BBa_I15009
 +
|BBa_I15010
 +
|}
 +
 +
[[Image:plategfp.jpg]]
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 +
*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.

Revision as of 17:46, 27 May 2008


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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
  • 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
  • While the following plates did not:
BBa_I14032 BBa_R0051 BBa_C0161
BBa_C0078 BBa_C0179 BBa_I15008
BBa_I15009 BBa_I15010

Plategfp.jpg

  • 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.
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