Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0032 mRFP Assay

From 2014.igem.org

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Transformation
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Double Transformation
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-dt(on pSB6A1)               each of 2.5&micro;L DNA to DH5&alpha;  
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-dt(on pSB6A1)
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each of 2.5&micro;L DNA to DH5&alpha;  
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-B0015(on pSB6A1)         LB 2 mL for 8hours Turbidity was controlled 0.1
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-B0015(on pSB6A1)    
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  LB 2 mL for 8hours  
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Turbidity was controlled 0.1
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Revision as of 02:42, 18 October 2014

Notebook
Lab Documents

140910 asB0034 & B0032 mRFP Assay

  • Start

  • Double Transformation

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-dt(on pSB6A1)
    each of 2.5µL DNA to DH5α
  • Colony PCR

    R0040-B0032-E1010-B0015(on pSB6A1) EcoRI_XbaI_F Tm:70.9℃ & RBS_SpeI_R Tm:59.2℃
  • Preculture

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0032-E1010-B0015(on pSB6A1)
    LB 2 mL for 8hours
    Turbidity was controlled 0.1
  • Culture

    M9ZB 2 mL IPTG 20 µ or not for 9 hours, 11 hours, 13hours
  • Assay

    fluoresence imageing 514 nm 610 filter
  • Complete!