Team:HokkaidoU Japan/Notebook/H Stem/Replacement To 1A3

From 2014.igem.org

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mini-Prep
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Mini-Prep
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      </ul>
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1A3 Vector
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R0010-stem-as-stem-B0015 (on pSB1A3)
 +
 
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      </ul>
      </ul>
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      <div class="step-recipe">
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Plac-stem-as-stem-dT on 1C3 with 100up_F & 200dn_R
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R0010-stem-as-stem-B0015(on pSB1C3) with 100up_F Tm:62.6&#8451; & 200dn_R Tm:66.6&#8451;
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      </ul>
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1A3 with EcoRI & PstI, Plac-stem-as-stem-dT with EcoRI & PstI
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pSB1A3 with EcoRI & PstI,  
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        R0010-stem-as-stem-B0015 with EcoRI & PstI
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      </ul>
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1A3 & Plac-stem-as-stem-dT
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0010-stem-as-stem-B0015 & pSB1A3
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      </ul>
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Plac-stem-as-stem-dT on 1A3
+
R0010-stem-as-stem-B0015(on pSB1A3)
 +
        5.0&micro;L to DH5alpha;
 +
 
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Revision as of 01:37, 18 October 2014

Notebook
Lab Documents

H-Stem Vector Exchange to pSB1A3

  • Start

  • Mini-Prep

    R0010-stem-as-stem-B0015 (on pSB1A3)
  • PCR

    R0010-stem-as-stem-B0015(on pSB1C3) with 100up_F Tm:62.6℃ & 200dn_R Tm:66.6℃
  • Digestion

    pSB1A3 with EcoRI & PstI, R0010-stem-as-stem-B0015 with EcoRI & PstI
  • Ligation

    0010-stem-as-stem-B0015 & pSB1A3
  • Transformation

    R0010-stem-as-stem-B0015(on pSB1A3) 5.0µL to DH5alpha;
  • Complete!