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Team:HokkaidoU Japan/Notebook/September1
From 2010.igem.org
(Difference between revisions)
(→PCR of 1-5A（RFP reporter）) |
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| 1 uL | | 1 uL | ||
|- | |- | ||
| - | |style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/ | + | |style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-5A]]) : |
| 1 uL | | 1 uL | ||
|- | |- | ||
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* 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: [[Team:HokkaidoU_Japan/ | + | * 5: [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-5A]] (1000bp): 80 ng/uL |
Revision as of 11:31, 16 October 2010
since 13 Jul 2010
since 1 Aug 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
