Team:HokkaidoU Japan/Notebook/plastic Week 9

From 2012.igem.org

(Difference between revisions)
Line 104: Line 104:
 +
 +
==Gel extraction==
 +
 +
Gel extraction for digestion product. Used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.
 +
 +
</div></div>
 +
 +
<div class="hokkaidou-notebook-daily">
 +
==July 3rd==
 +
<div>
 +
==Digestion==
 +
<p>
 +
Digestion to divide PhaA, PhaB and PhaC(BBa_K342001) with XbaI and PstI.<br />And PhaC(BBa_K342001) and pSB1C3 with XbaI and SpeI.
 +
<br /><br />
 +
PhaA
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (125 ng/ul)
 +
  |6.6 ul
 +
  |-
 +
  |XbaI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xM buffer
 +
  |2 ul
 +
  |-
 +
  |DW
 +
  |9.4 ul
 +
  |-
 +
  |Total
 +
  |20 ul
 +
  |}
 +
<br />
 +
PhaB
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (125 ng/ul)
 +
  |4 ul
 +
  |-
 +
  |XbaI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xM buffer
 +
  |2 ul
 +
  |-
 +
  |DW
 +
  |12 ul
 +
  |-
 +
  |Total
 +
  |20 ul
 +
  |}
 +
<br />
 +
 +
PhaC(BBa_K342001)
 +
{|class="hokkaidou-table-digestion"
 +
  |-
 +
  |DNA solution (125 ng/ul)
 +
  |10 ul
 +
  |-
 +
  |XbaI
 +
  |1 ul
 +
  |-
 +
  |PstI
 +
  |1 ul
 +
  |-
 +
  |10xM buffer
 +
  |2 ul
 +
  |-
 +
  |DW
 +
  |0.9 ul
 +
  |-
 +
  |Total
 +
  |20 ul
 +
  |}
 +
<br />
 +
 +
</p>
==Gel extraction==
==Gel extraction==

Revision as of 11:50, 28 August 2012

Contents

July 2nd

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.

July 3rd

Digestion

Digestion to divide PhaA, PhaB and PhaC(BBa_K342001) with XbaI and PstI.
And PhaC(BBa_K342001) and pSB1C3 with XbaI and SpeI.

PhaA

DNA solution (125 ng/ul) 6.6 ul
XbaI 1 ul
PstI 1 ul
10xM buffer 2 ul
DW 9.4 ul
Total 20 ul


PhaB

DNA solution (125 ng/ul) 4 ul
XbaI 1 ul
PstI 1 ul
10xM buffer 2 ul
DW 12 ul
Total 20 ul


PhaC(BBa_K342001)

DNA solution (125 ng/ul) 10 ul
XbaI 1 ul
PstI 1 ul
10xM buffer 2 ul
DW 0.9 ul
Total 20 ul


Gel extraction

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



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