"
Team:HokkaidoU Japan/Notebook/asB0034/140910 asB0034&B0034 GFP Assay
From 2014.igem.org
(Difference between revisions)
| Line 130: | Line 130: | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min | Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min | ||
| - | each of 2.5µL DNA to DH5α | + | each of 2.5µL DNA to DH5α 42℃ 30sec |
on ice 2min | on ice 2min | ||
</div> | </div> | ||
| Line 153: | Line 153: | ||
<div class="step-recipe"> | <div class="step-recipe"> | ||
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) | Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) | ||
| - | pHN1257_up_F Tm:60.1 | + | pHN1257_up_F Tm:60.1℃ |
| - | asB0034-R Tm:68.7 | + | asB0034-R Tm:68.7℃ |
</div> | </div> | ||
</div> | </div> | ||
Revision as of 14:43, 17 October 2014
hogehoge
140910 asB0034 & B0034 GFP Assay
-
Start
-
Transformation
Ptrc-stem-asB0034-stem on pHN1257&Ptet-B0034-GFP-dt on pSB6A1 on ice 30min each of 2.5µL DNA to DH5α 42℃ 30sec on ice 2min -
Colony PCR
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-mRFP-dT(pSB6A1) pHN1257_up_F Tm:60.1℃ asB0034-R Tm:68.7℃ -
Preculture
Ptrc-stem-asB0034-stem(pHN1257)&Ptet-B0034-GFP-dT(pSB6A1) LB 2 mL for 9hours Turbidity was controlled 0.1 -
Culture
M9ZB 2 mL IPTG 20 µ or not for 9 hours, 11 hours, 13hours -
Assay
fluoresence imageing 488 nm 530 filter -
Complete!
