Team:HokkaidoU Japan/Notebook/September1
From 2010.igem.org
(Difference between revisions)
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- | =1-5A(RFP | + | =PCR of 1-5A(RFP reporter)= |
{|class="protocol" | {|class="protocol" | ||
|style="text-align:right;"|'''Reagent''' | |style="text-align:right;"|'''Reagent''' | ||
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|} | |} | ||
- | * | + | * Because part lenght was 1069bp plus primer added 300bp to that, the extension time was 90 sec |
- | * | + | * PCR product was purified via Microcon YM-10 |
- | === | + | |
- | [[Image:HokkaidoU Japan 20100901a.JPG|200px|right|thumb|]] | + | ===Electrophoresis=== |
- | pUC119,pSB1C3,1- | + | |
- | * 2: | + | [[Image:HokkaidoU Japan 20100901a.JPG|200px|right|thumb|Electrophoresis of purified PCR product]] |
+ | |||
+ | Electrophoresed pUC119,pSB1C3,1-5A to check concentration (1 uL of each) | ||
+ | * 2: [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''d III](2 uL) | ||
* 3: pUC119 (3000bp): 10 ng/uL | * 3: pUC119 (3000bp): 10 ng/uL | ||
* 4: pSB1C3 (2000bp): 20 ng/uL | * 4: pSB1C3 (2000bp): 20 ng/uL | ||
* 5: 1-5A (1000bp): 80 ng/uL | * 5: 1-5A (1000bp): 80 ng/uL |
Revision as of 17:13, 27 September 2010
PCR of 1-5A(RFP reporter)
Reagent | Amount |
Autoclaved DW : | 33 uL |
10x PCR buffer : | 5 uL |
2 mM dNTPs : | 5 uL |
25 mM MgSO4 : | 3 uL |
100 bp up primer : | 1 uL |
200 bp down primer : | 1 uL |
KOD plus Neo : | 1 uL |
Template (1-5A) : | 1 uL |
Total : | 50 uL |
- Because part lenght was 1069bp plus primer added 300bp to that, the extension time was 90 sec
- PCR product was purified via Microcon YM-10
Electrophoresis
Electrophoresed pUC119,pSB1C3,1-5A to check concentration (1 uL of each)
- 2: Lambda/Hind III(2 uL)
- 3: pUC119 (3000bp): 10 ng/uL
- 4: pSB1C3 (2000bp): 20 ng/uL
- 5: 1-5A (1000bp): 80 ng/uL