Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 mRFP Assay

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Transformation
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Double Transformation
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Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-mRFP-dt on pSB6A1 on ice 30min each of 2.5&micro;L DNA to DH5&alpha; 42&#8451;    30sec on ice  2min
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1)  
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each of 2.5&micro;L to DH5&alpha;
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1)
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Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1) with pHN1257_up_F Tm:60.1&#8451; & asB0034_R Tm:68.7&#8451;
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pHN1257_up_F Tm:60.1&#8451;
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asB0034-R Tm:68.7&#8451;
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Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1)
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Ptrc-stem-asB0034-stem(pHN1257) & R0040-B0034-E1010-B0015(pSB6A1)
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LB 2 mL
LB 2 mL
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for 3hours
for 3hours
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Turbidity was controlled 0.1
Turbidity was controlled 0.1
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LB 2 mL
LB 2 mL
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IPTG 20 &micro; or not
IPTG 20 &micro; or not
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for 11 hours, 13hours, 15hours, 17 hours
for 11 hours, 13hours, 15hours, 17 hours
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Latest revision as of 15:31, 9 September 2015

Notebook
Lab Documents

140910 asB0034&B0034 mRFP Assay

  • Start

  • Double Transformation

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1)
    each of 2.5µL to DH5α

  • Colony PCR

    Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1) with pHN1257_up_F Tm:60.1℃ & asB0034_R Tm:68.7℃

  • Preculture

    Ptrc-stem-asB0034-stem(pHN1257) & R0040-B0034-E1010-B0015(pSB6A1)
    LB 2 mL
    for 3hours
    Turbidity was controlled 0.1

  • Culture

    LB 2 mL
    IPTG 20 µ or not
    for 11 hours, 13hours, 15hours, 17 hours

  • Assay

    fluoresence imageing 514 nm 610 filter

  • Complete!

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