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Team:HokkaidoU Japan/Notebook/asB0034/asB0034-construction
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| - | + | Ptrc-stem-asB0034-stem on pHN1257 | |
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| + | 5.0µL DNA to DH5α | ||
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| - | Ptrc-stem-asB0034-stem(on pHN1257)&R0040-B0034-E1010-B0015(on pSB6A1) | + | Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1) |
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each of 2.5µL DNA to DH5α | each of 2.5µL DNA to DH5α | ||
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Latest revision as of 15:24, 9 September 2015
Notebook
Lab Documents
asB0034 Construction
-
Start
-
Primer Annealing
asB0034_F Tm:68.7℃ asB0034_R Tm:68.7℃ -
Digestion
Cut pHN1257 with NcoI, XhoI(using 10×cut smart) -
Gel extraction
digested pHN1257 -
Ethanol Precipetation
digested pHN1257 -
Ligation
Ligate pHN1257 with asB0034 -
Transformation
Ptrc-stem-asB0034-stem on pHN12575.0µL DNA to DH5αColony PCR
Ptrc-stem-asB0034-stem(on pHN1257) pHN1257_up_F Tm:60.1℃ & asB0034-R Tm:68.7℃Culture
Ptrc-stem-asB0034-stem(on pHN1257)Mini-Prep
Ptrc-stem-asB0034-stem(on pHN1257)Double Transformation
Ptrc-stem-asB0034-stem(on pHN1257) & R0040-B0034-E1010-B0015(on pSB6A1)each of 2.5µL DNA to DH5αComplete!
