Team:Cambridge/Notebook/12

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(New page: <div align="center">Week 11: Monday 27th September - Sunday 3rd October</div> µl°C)
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<div align="center">Week 11: Monday 27th September - Sunday 3rd October</div>
<div align="center">Week 11: Monday 27th September - Sunday 3rd October</div>
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===113. Expt: Biobrick assembly of fluorescent proteins (YFP, CFP, RFP) into PP+pBAD+pSB1C3 and pBAD+pSB1C3 (Emily and Bill)===
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*Miniprepped overnight cultures of YFP, CFP, RFP with rbs's and PP+pBAD+pSB1C3
 +
*Nanodrop readings:
 +
{|class="wikitable"
 +
|-
 +
|
 +
|Nanodrop reading (ng/µl)
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|-
 +
|Cm+PP+pBAD+pSB1C3
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|48.5
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|-
 +
|YFP+rbs
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|74.8
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|-
 +
|RFP+rbs
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|32.9
 +
|-
 +
|CFP+rbs
 +
|46.7
 +
|}
 +
 +
pSB1C3 from freezer was used at 12.9ng/µl
 +
 +
*Restrict using protocol on p88, using following quantities:
 +
{|class="wikitable"
 +
|-
 +
|
 +
|Cm+PP+pSB1C3
 +
|YFP (for PP)
 +
|YFP (for pBAD)
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|RFP (for PP)
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|RFP (for pBAD)
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|CFP (for PP)
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|CFP (for pBAD)
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|pSB1C3
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|-
 +
|Nuclease-free H20
 +
|1
 +
|6
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|6
 +
|1
 +
|1
 +
|6
 +
|6
 +
|1
 +
|-
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|10x FD Buffer
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|2
 +
|2
 +
|2
 +
|2
 +
|2
 +
|2
 +
|2
 +
|2
 +
|-
 +
|Plasmid DNA
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|15
 +
|10
 +
|10
 +
|15
 +
|15
 +
|10
 +
|10
 +
|16
 +
|-
 +
|EcoRI
 +
|0
 +
|0
 +
|0
 +
|0
 +
|0
 +
|0
 +
|0
 +
|1
 +
|-
 +
|SpeI
 +
|0
 +
|1
 +
|0
 +
|1
 +
|0
 +
|1
 +
|0
 +
|0
 +
|-
 +
|XbaI
 +
|1
 +
|0
 +
|1
 +
|0
 +
|1
 +
|0
 +
|1
 +
|0
 +
|-
 +
|PstI
 +
|1
 +
|1
 +
|1
 +
|1
 +
|1
 +
|1
 +
|1
 +
|1
 +
|-
 +
|
 +
|*
 +
|*
 +
|
 +
|*
 +
|
 +
|*
 +
|
 +
|
 +
|}
µl°C
µl°C

Revision as of 11:21, 7 October 2010

Week 11: Monday 27th September - Sunday 3rd October

113. Expt: Biobrick assembly of fluorescent proteins (YFP, CFP, RFP) into PP+pBAD+pSB1C3 and pBAD+pSB1C3 (Emily and Bill)

  • Miniprepped overnight cultures of YFP, CFP, RFP with rbs's and PP+pBAD+pSB1C3
  • Nanodrop readings:
Nanodrop reading (ng/µl)
Cm+PP+pBAD+pSB1C3 48.5
YFP+rbs 74.8
RFP+rbs 32.9
CFP+rbs 46.7

pSB1C3 from freezer was used at 12.9ng/µl

  • Restrict using protocol on p88, using following quantities:
Cm+PP+pSB1C3 YFP (for PP) YFP (for pBAD) RFP (for PP) RFP (for pBAD) CFP (for PP) CFP (for pBAD) pSB1C3
Nuclease-free H20 1 6 6 1 1 6 6 1
10x FD Buffer 2 2 2 2 2 2 2 2
Plasmid DNA 15 10 10 15 15 10 10 16
EcoRI 0 0 0 0 0 0 0 1
SpeI 0 1 0 1 0 1 0 0
XbaI 1 0 1 0 1 0 1 0
PstI 1 1 1 1 1 1 1 1
* * * *

µl°C

Retrieved from "http://2010.igem.org/Team:Cambridge/Notebook/12"