"
Team:HokkaidoU Japan/Notebook/September1
From 2010.igem.org
(Difference between revisions)
| Line 27: | Line 27: | ||
| 1 uL | | 1 uL | ||
|- | |- | ||
| - | |style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/ | + | |style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/Parts#BioBricks|1-5A]]) : |
| 1 uL | | 1 uL | ||
|- | |- | ||
| Line 34: | Line 34: | ||
|} | |} | ||
| - | * Because part lenght was | + | * Because part lenght was 1069 bp plus primer added 300 bp to that, the extension time was 90 sec |
* PCR product was purified via Microcon YM-10 | * PCR product was purified via Microcon YM-10 | ||
| Line 42: | Line 42: | ||
Electrophoresed pUC119，pSB1C3，1-5A to check concentration （1 uL of each） | Electrophoresed pUC119，pSB1C3，1-5A to check concentration （1 uL of each） | ||
| - | * 2: [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png | + | * 2: [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''d III]（2 uL) |
| - | * 3: pUC119 ( | + | * 3: pUC119 (3000 bp): 10 ng/uL |
| - | * 4: pSB1C3 ( | + | * 4: pSB1C3 (2000 bp): 20 ng/uL |
| - | * 5: [[Team:HokkaidoU_Japan/ | + | * 5: [[Team:HokkaidoU_Japan/Parts#BioBricks|1-5A]] (1000 bp): 80 ng/uL |
Latest revision as of 08:01, 27 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 1069 bp plus primer added 300 bp 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: λ/Hind III（2 uL)
- 3: pUC119 (3000 bp): 10 ng/uL
- 4: pSB1C3 (2000 bp): 20 ng/uL
- 5: 1-5A (1000 bp): 80 ng/uL
