Team:HokkaidoU Japan/Notebook/September21
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For this I ordered oligo primers for sigma | For this I ordered oligo primers for sigma | ||
<br><br><br><br><br> | <br><br><br><br><br> | ||
- | == | + | == Arabinose induced expression of GFP == |
- | + | ||
*GFP-doubleterminator | *GFP-doubleterminator | ||
BBa_I13401 1-12K 875bp pSB1A2 | BBa_I13401 1-12K 875bp pSB1A2 | ||
- | * | + | *Arabinose promoter |
BBa_I0050 3-20B 1210bp pSB2K3 | BBa_I0050 3-20B 1210bp pSB2K3 | ||
*pSB1C3 2072bp | *pSB1C3 2072bp | ||
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<br> | <br> | ||
- | 1) | + | 1)PCR mix of Arabinose promoter and GFP, pSB1A3 |
{|style="text-align:center; margin-left:100px;" class="protocol" | {|style="text-align:center; margin-left:100px;" class="protocol" | ||
!Reagent | !Reagent | ||
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2) | 2) | ||
+ | * 94C 2min | ||
+ | * 98C 10sec | ||
+ | * 68C 2min | ||
+ | ** 35cycle | ||
+ | * 4C hold | ||
- | + | 3)After PCR, Electrophored and extracted samples | |
- | + | 4)Estimated the concentration of the samples | |
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- | 4) | + | |
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<div style="margin-left:100px;"> | <div style="margin-left:100px;"> | ||
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- | + | from left to right | |
- | 1.λ/Hind | + | 1.λ/Hind III, EcoR I |
- | 2. | + | 2.Arabinose promoter |
3.GFP+double terminator | 3.GFP+double terminator |
Latest revision as of 02:35, 28 October 2010
Annealing of RBS made from oligos
Annealing of RBS (BBa_B0034) made from oligos
For this I ordered oligo primers for sigma
Arabinose induced expression of GFP
- GFP-doubleterminator
BBa_I13401 1-12K 875bp pSB1A2
- Arabinose promoter
BBa_I0050 3-20B 1210bp pSB2K3
- pSB1C3 2072bp
- pSB1A3 2157bp
1)PCR mix of Arabinose promoter and GFP, pSB1A3
Reagent | Amount |
---|---|
Dw | 33 uL |
10×PCR buffur | 5 |
2mM 4dNTP | 5 |
MgSO4 | 3 |
Ex-F | 1 |
Ps-R | 1 |
KOD neo plus | 1 |
DNA | 1 |
Total | 50 uL |
2)
- 94C 2min
- 98C 10sec
- 68C 2min
- 35cycle
- 4C hold
3)After PCR, Electrophored and extracted samples
4)Estimated the concentration of the samples