Team:HokkaidoU Japan/Notebook/September1

From 2010.igem.org

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|style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-5A]]) :  
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|style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/Parts#BioBricks|1-5A]]) :  
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* Because part lenght was 1069bp plus primer added 300bp to that, the extension time was 90 sec
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* 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
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Electrophoresed pUC119,pSB1C3,1-5A to check concentration (1 uL of each)
Electrophoresed pUC119,pSB1C3,1-5A to check concentration (1 uL of each)
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* 2: [http://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''d III](2 uL)
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* 2: [http://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''d III](2 uL)
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* 3: pUC119 (3000bp): 10 ng/uL
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* 3: pUC119 (3000 bp): 10 ng/uL
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* 4: pSB1C3 (2000bp): 20 ng/uL
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* 4: pSB1C3 (2000 bp): 20 ng/uL
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* 5: [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-5A]] (1000bp): 80 ng/uL
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* 5: [[Team:HokkaidoU_Japan/Parts#BioBricks|1-5A]] (1000 bp): 80 ng/uL

Latest revision as of 08:01, 27 October 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

Electrophoresis of purified PCR product

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