Team:HokkaidoU Japan/Notebook/September22

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{{Template:HokkaidoU_Japan}}
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= Digestion of pSB1A3, Arabinose Promoter & GFP + double terminator =
 +
 +
*Digestion mix -> XbaI in GFP + double terminator solution required putting on incubator for a long time, so did it.
 +
 +
{|style="text-align:center; float:left;" class="protocol"
 +
!Reagent
 +
!Amount
 +
|-
 +
|10x H buffer
 +
|2 uL
 +
|-
 +
|DW
 +
|10
 +
|-
 +
|pSB1A3
 +
|15
 +
|-
 +
|0.1% BSA
 +
|2
 +
|-
 +
|EcoR I
 +
|0.5
 +
|-
 +
|Pst I
 +
|0.5
 +
|-
 +
|style="border-top:1px solid #996;"|'''Total'''
 +
|style="border-top:1px solid #996;"|'''30 uL'''
 +
|}
 +
{|style="text-align:center; float:left;" class="protocol"
 +
!Reagent
 +
!Amount
 +
|-
 +
|10x H buffer
 +
|2 uL
 +
|-
 +
|DW
 +
|8
 +
|-
 +
|Arabinose Promoter
 +
|6.3
 +
|-
 +
|0.1% BSA
 +
|2
 +
|-
 +
|EcoR I
 +
|1
 +
|-
 +
|Spe I
 +
|0.7
 +
|-
 +
|style="border-top:1px solid #996;"|'''Total'''
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|style="border-top:1px solid #996;"|'''20 uL'''
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|}
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{|style="text-align:center; float:left;" class="protocol"
 +
!Reagent
 +
!Amount
 +
|-
 +
|10x M buffer
 +
|5 uL
 +
|-
 +
|DW
 +
|32.5
 +
|-
 +
|GFP + double terminator
 +
|3
 +
|-
 +
|BSA
 +
|5
 +
|-
 +
|Xba I
 +
|4
 +
|-
 +
|Pst I
 +
|0.5
 +
|-
 +
|style="border-top:1px solid #996;"|'''Total'''
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|style="border-top:1px solid #996;"|'''50 uL'''
 +
|}
 +
<div style="clear:both"></div>
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 +
*Precipitated two samples by EtOH after extracted them from a gel.
 +
->Ligation & Transformation

Latest revision as of 17:57, 27 October 2010

Notebook

Digestion of pSB1A3, Arabinose Promoter & GFP + double terminator

  • Digestion mix -> XbaI in GFP + double terminator solution required putting on incubator for a long time, so did it.
Reagent Amount
10x H buffer 2 uL
DW 10
pSB1A3 15
0.1% BSA 2
EcoR I 0.5
Pst I 0.5
Total 30 uL
Reagent Amount
10x H buffer 2 uL
DW 8
Arabinose Promoter 6.3
0.1% BSA 2
EcoR I 1
Spe I 0.7
Total 20 uL
Reagent Amount
10x M buffer 5 uL
DW 32.5
GFP + double terminator 3
BSA 5
Xba I 4
Pst I 0.5
Total 50 uL
  • Precipitated two samples by EtOH after extracted them from a gel.
->Ligation & Transformation
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