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

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mini-Prep
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Mini-Prep
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1A3 Vector
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R0010-stem-as-stem-B0015 (on pSB1A3)
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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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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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1A3 & Plac-stem-as-stem-dT
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Ligate 0010-stem-as-stem-B0015 with pSB1A3
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Plac-stem-as-stem-dT on 1A3
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R0010-stem-as-stem-B0015(on pSB1A3)
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5.0&micro;L to DH5alpha;
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Latest revision as of 15:28, 9 September 2015

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

    Ligate 0010-stem-as-stem-B0015 with pSB1A3

  • Transformation

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

  • Complete!

Retrieved from "http://2014.igem.org/Team:HokkaidoU_Japan/Notebook/H_Stem/Replacement_To_1A3"