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Team:Bordeaux/NotebookJune12
From 2014.igem.org
(Difference between revisions)
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<br><br> | <br><br> | ||
| - | <div class="type-exp">Production</div> | + | <div class="type-exp">Production</div><br> |
<div style="margin-left:65px"> | <div style="margin-left:65px"> | ||
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A<sub>600</sub> preculture VPGXG-40 = 2,431 | A<sub>600</sub> preculture VPGXG-40 = 2,431 | ||
A<sub>600</sub> preculture VPGXG-60 = 2,143 | A<sub>600</sub> preculture VPGXG-60 = 2,143 | ||
| - | <div style="margin-left:85px"> - Culture of VPGXG-20, VPGXG-40 and VPGXG-60 at 37°C </div><br> | + | <div style="margin-left:85px"> - Culture of VPGXG-20, VPGXG-40 and VPGXG-60 at 37°C </div><br><br> |
T=0, A<sub>600</sub> VPGXG-20 = 0,003 | T=0, A<sub>600</sub> VPGXG-20 = 0,003 | ||
A<sub>600</sub> VPGXG-40 = 0,005 | A<sub>600</sub> VPGXG-40 = 0,005 | ||
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<br><br> | <br><br> | ||
| - | <div class="type-exp">Purification</div> | + | <div class="type-exp">Purification</div><br> |
| + | |||
<ul class="day-detail"> | <ul class="day-detail"> | ||
<li>We stop protocol after resuspend cell pellet in 20mL PBS and store at -80°C.</li><br> | <li>We stop protocol after resuspend cell pellet in 20mL PBS and store at -80°C.</li><br> | ||
Revision as of 21:43, 17 October 2014
6th June
Molecular biology
- Making competent cell: day 3
- Digestion, ligation and transformation of pSB1C3 and pSB6A1 with our inserts VPGXG-20, VPGXG-40 and VPGXG-60.
Production
A600 preculture VPGXG-20 = 2,349
A600 preculture VPGXG-40 = 2,431
A600 preculture VPGXG-60 = 2,143
T=0, A600 VPGXG-20 = 0,003 A600 VPGXG-40 = 0,005 A600 VPGXG-60 = 0,047
- Culture of VPGXG-20, VPGXG-40 and VPGXG-60 at 37°C
T=0, A600 VPGXG-20 = 0,003 A600 VPGXG-40 = 0,005 A600 VPGXG-60 = 0,047
- IPTG induction when A600 = 0,8 and culture at 25°C
Purification
- We stop protocol after resuspend cell pellet in 20mL PBS and store at -80°C.
