Exeter/29 August 2013

From 2013.igem.org

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{{:Team:Exeter/Template/Header}}
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<div class="container">
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    <div class="row">
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        <div class="span" style="text-align:justify">
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== Minipreps ==
== Minipreps ==
Line 5: Line 10:
* 1 + 4
* 1 + 4
-
* K592011 B
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* [http://parts.igem.org/Part:BBa_K592011 K592011] B
-
* K864404
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* [http://parts.igem.org/Part:BBa_K864404 K864404]
* Cph8 #3
* Cph8 #3
Line 13: Line 18:
* 2 + 4
* 2 + 4
-
* K592022 B
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* [http://parts.igem.org/Part:BBa_K592022 K592022] B
* ENvZ no antibiotic
* ENvZ no antibiotic
Line 22: Line 27:
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The liquid cultures of K592011 A and K592022 A did not grow.
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The liquid cultures of [http://parts.igem.org/Part:BBa_K592011 K592011] A and [http://parts.igem.org/Part:BBa_K592022 K592022] A did not grow.
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==NanoDrop==
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 +
The NanoDrop machine was zero-ed using the elution buffer from the gel extraction kit used.
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*K592011 B - 164.2 ng/ul
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*B0015 - 37.7 ng/ul
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*OmpR prom - 50.7 ng/ul
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*RBS - 49.1 ng/ul
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 +
Two different RFP controls were nanodropped
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*RFP control - 61.7 ng/ul
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*RFP control - 75.9 ng/ul
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The first RFP control was used in the digestion of K592011 ans B0015.
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The second RFP control was used in the digestion of OmpR and RBS.
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==Digestion==
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== NanoDrop from miniprep ==
 
{| class="wikitable"
{| class="wikitable"
|-
|-
-
! Part !! Concentration (ng/ul)
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! Part !! K592011 (Part A) !! B0015 (Part B) !! AMP Plasmid !! RFP (Control)
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|-
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| DNA || 1.52 ul || 6.63 ul || 10.0 ul || 4.05 ul
 +
|-
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| Non-nuclease Water || 14.48 ul || 9.37 ul || 5.5 ul || 11.95 ul
 +
|-
 +
| NEB Buffer || 2.5 ul || 2.5 ul || 2.5 ul || 2.5 ul
 +
|-
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| BSA || 0.5 ul || 0.5 ul || 0.5 ul || 0.5 ul
 +
|-
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| <i>EcoRI</i> || 0.5 ul || - || 0.5 ul || 0.5 ul
 +
|-
 +
| <i>SpeI</i> || 0.5 || - || - || -
 +
|-
 +
| <i>XbaI</i> || - || 0.5 ul || - || -
 +
|-
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| <i>PstI</i> || - || 0.5 ul || 0.5 ul || 0.5 ul
 +
|-
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| <i>DpnI</i> || - || - || 0.5 ul || -
 +
|}
 +
 
 +
 
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{| class="wikitable"
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|-
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! Part !! OmpR (Part A) !! RBS (Part B) !! AMP Plasmid !! RFP (Control)
 +
|-
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| DNA || 4.93 ul || 5.09 ul || 10.0 ul || 3.30 ul
 +
|-
 +
| Non-nuclease Water || 11.07 ul || 10.91 ul || 5.5 ul || 12.70 ul
 +
|-
 +
| NEB Buffer || 2.5 ul || 2.5 ul || 2.5 ul || 2.5 ul
 +
|-
 +
| BSA || 0.5 ul || 0.5 ul || 0.5 ul || 0.5 ul
 +
|-
 +
| <i>EcoRI</i> || 0.5 ul || - || 0.5 ul || 0.5 ul
|-
|-
-
| K592011 B ||164.2
+
| <i>SpeI</i> || 0.5 || - || - || -
|-
|-
-
| B0015 || 37.7
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| <i>XbaI</i> || - || 0.5 ul || - || -
|-
|-
-
| OmpR promoter || 50.7
+
| <i>PstI</i> || - || 0.5 ul || 0.5 ul || 0.5 ul
|-
|-
-
| RBS || 49.1
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| <i>DpnI</i> || - || - || 0.5 ul || -
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|}
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Take me back to the [https://2013.igem.org/Team:Exeter/Notebook notebook].
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== Digestion ==
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  </div>
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</div>
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{{:Team:Exeter/Template/Footer}}

Latest revision as of 20:37, 2 October 2013

Exeter iGEM 2013 · Paint by Coli

Minipreps

We miniprepped:

  • 1 + 4
  • Cph8 #3
  • 2 + 4
  • ENvZ no antibiotic
  • ENvZ
  • Cph8 #5


The liquid cultures of K592011 A and K592022 A did not grow.

NanoDrop

The NanoDrop machine was zero-ed using the elution buffer from the gel extraction kit used.

  • K592011 B - 164.2 ng/ul
  • B0015 - 37.7 ng/ul
  • OmpR prom - 50.7 ng/ul
  • RBS - 49.1 ng/ul

Two different RFP controls were nanodropped

  • RFP control - 61.7 ng/ul
  • RFP control - 75.9 ng/ul

The first RFP control was used in the digestion of K592011 ans B0015. The second RFP control was used in the digestion of OmpR and RBS.


Digestion

Part K592011 (Part A) B0015 (Part B) AMP Plasmid RFP (Control)
DNA 1.52 ul 6.63 ul 10.0 ul 4.05 ul
Non-nuclease Water 14.48 ul 9.37 ul 5.5 ul 11.95 ul
NEB Buffer 2.5 ul 2.5 ul 2.5 ul 2.5 ul
BSA 0.5 ul 0.5 ul 0.5 ul 0.5 ul
EcoRI 0.5 ul - 0.5 ul 0.5 ul
SpeI 0.5 - - -
XbaI - 0.5 ul - -
PstI - 0.5 ul 0.5 ul 0.5 ul
DpnI - - 0.5 ul -


Part OmpR (Part A) RBS (Part B) AMP Plasmid RFP (Control)
DNA 4.93 ul 5.09 ul 10.0 ul 3.30 ul
Non-nuclease Water 11.07 ul 10.91 ul 5.5 ul 12.70 ul
NEB Buffer 2.5 ul 2.5 ul 2.5 ul 2.5 ul
BSA 0.5 ul 0.5 ul 0.5 ul 0.5 ul
EcoRI 0.5 ul - 0.5 ul 0.5 ul
SpeI 0.5 - - -
XbaI - 0.5 ul - -
PstI - 0.5 ul 0.5 ul 0.5 ul
DpnI - - 0.5 ul -

Take me back to the notebook.

Exeter iGEM 2013 · Paint by Coli
Retrieved from "http://2013.igem.org/Exeter/29_August_2013"