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Team:HokkaidoU Japan/Notebook/asB0034/asB0034-construction
From 2014.igem.org
(Difference between revisions)
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</ul> | </ul> | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
| - | Ptrc-stem-asB0034-stem on pHN1257 | + | Ptrc-stem-asB0034-stem on pHN1257 |
5.0µL DNA to DH5α | 5.0µL DNA to DH5α | ||
| Line 329: | Line 329: | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
Ptrc-stem-asB0034-stem(on pHN1257)&R0040-B0034-E1010-B0015(on pSB6A1) | Ptrc-stem-asB0034-stem(on pHN1257)&R0040-B0034-E1010-B0015(on pSB6A1) | ||
| - | + | each of 2.5µL DNA to DH5α | |
</div> | </div> | ||
</div> | </div> | ||
Revision as of 00:35, 18 October 2014
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 pHN1257 5.0µL 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!
