Team:HokkaidoU Japan/Notebook/plastic Week 9

From 2012.igem.org

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==Digestion==
 +
<p>
 +
Digestion to divide BBa_K342001(PhaC) with XbaI and PstI.<br />And BBa_B0034(RBS) with SpeI and PstI (with three samples).
 +
<br /><br />
 +
PhaC (BBa_K342001)
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (100 ng/ul)
 +
  |12.5 ul
 +
  |-
 +
  |XbaI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xM buffer
 +
  |2 ul
 +
  |-
 +
  |DW
 +
  |3.5 ul
 +
  |-
 +
  |Total
 +
  |20 ul
 +
  |}
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<br />
 +
RBS (BBa_B0034)
 +
<br />
 +
N0.1
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (20.3 ng/ul)
 +
  |14.3 ul
 +
  |-
 +
  |SpeI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xH buffer
 +
  |2.5 ul
 +
  |-
 +
  |DW
 +
  |0.2 ul
 +
  |-
 +
  |Total
 +
  |25 ul
 +
  |}
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<br /><br />
 +
N0.2
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (15.6 ng/ul)
 +
  |18.6 ul
 +
  |-
 +
  |SpeI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xH buffer
 +
  |2.5 ul
 +
  |-
 +
  |DW
 +
  |1.9 ul
 +
  |-
 +
  |Total
 +
  |25 ul
 +
  |}
 +
<br />
 +
 +
N0.3<br />
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (16.9 ng/ul)
 +
  |17.2 ul
 +
  |-
 +
  |SpeI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xH buffer
 +
  |2.5 ul
 +
  |-
 +
  |DW
 +
  |3.3 ul
 +
  |-
 +
  |Total
 +
  |25 ul
 +
  |}
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</p>
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 +
</div></div>
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 +
==Gel extraction==
 +
<p>
 +
Gel extraction for digestion product. Used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.
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</p>
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</div></div>
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 +
<div class="hokkaidou-notebook-daily">
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Revision as of 13:17, 27 August 2012

Digestion

Digestion to divide BBa_K342001(PhaC) with XbaI and PstI.
And BBa_B0034(RBS) with SpeI and PstI (with three samples).

PhaC (BBa_K342001)

DNA solution (100 ng/ul) 12.5 ul
XbaI 1 ul
PstI 1 ul
10xM buffer 2 ul
DW 3.5 ul
Total 20 ul


RBS (BBa_B0034)
N0.1

DNA solution (20.3 ng/ul) 14.3 ul
SpeI 1 ul
PstI 1 ul
10xH buffer 2.5 ul
DW 0.2 ul
Total 25 ul



N0.2

DNA solution (15.6 ng/ul) 18.6 ul
SpeI 1 ul
PstI 1 ul
10xH buffer 2.5 ul
DW 1.9 ul
Total 25 ul


N0.3

DNA solution (16.9 ng/ul) 17.2 ul
SpeI 1 ul
PstI 1 ul
10xH buffer 2.5 ul
DW 3.3 ul
Total 25 ul

Gel extraction

Gel extraction for digestion product. Used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.


</div> </div> </div>

Retrieved from "http://2012.igem.org/Team:HokkaidoU_Japan/Notebook/plastic_Week_9"