NYMU Taipei/Lab Notes/2007 9 29

From 2007.igem.org

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<p>
PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS
PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS
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</p>
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<br><br>
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<p>
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PCR for IDE
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{| align=left | border=1 | style="text-align:center"
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!
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!size
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!Primer
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!Template
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|-
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|IDE
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|140bp
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|
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|M_IDE
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|-
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|}
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{| align=right | border=1 | style="text-align:center"
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|Template
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|1uL
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|-
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|10X buffer
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|5uL
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|-
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|2.5mM dNTP
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|2uL
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|-
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|Primer_F
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|1.5uL
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|-
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|Primer_R
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|1.5uL
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|-
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|pfu polymerase
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|0.5uL
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|-
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|ddH2O
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|38.5uL
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|-
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|Total
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|50uL
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|-}
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{| align=center | border=1 | style="text-align:center"
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!Step
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!Temp (°C)
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!Time (mins)
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!Cycle
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|-
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|Initial Denaturation
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|95
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|10
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|
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|-
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|Denatuation<br>Annealing<br>Extension
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|95<br>55<br>72
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|1<br>1<br>4
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|35
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|-
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 +
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|Final Extension
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|72
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|10
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|
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|-
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|Soak
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|4
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|
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|
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|-
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|}
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</p>

Revision as of 12:59, 29 September 2007

PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS

size Primer Template
1 TAT_A 140bp TAT_F+TAT_A_R E.coli K12 genome DNA
2 INS_A 111bp INS_A_F+INS_A_R MGC clone (H_INS 1)
3 TAT_B 137bp TAT_F+TAT_B_R E.coli K12 genome DNA
4 INS_B 135bp INS_B_F+INS_B_R MGC clone (H_INS 1)
5 TAT_D 131bp TAT_F+TAT_D_R E.coli K12 genome DNA
6 OmpRBS 100bp OmpRBS_F+OmpRBS_R pOmpC
Template 1uL
10X buffer 5uL
2.5mM dNTP 2uL
Primer_F 1.5uL
Primer_R 1.5uL
pfu polymerase 0.5uL
ddH2O 38.5uL
Total 50uL
Step Temp (°C) Time (mins) Cycle
Initial Denaturation 95 10
Denatuation
Annealing
Extension 		95
55
72 		1
1
1 		35
Final Extension 72 10
Soak 4



PCR for IDE

size Primer Template
IDE 140bp M_IDE
Template 1uL
10X buffer 5uL
2.5mM dNTP 2uL
Primer_F 1.5uL
Primer_R 1.5uL
pfu polymerase 0.5uL
ddH2O 38.5uL
Total 50uL
Step Temp (°C) Time (mins) Cycle
Initial Denaturation 95 10
Denatuation
Annealing
Extension 		95
55
72 		1
1
4 		35
Final Extension 72 10
Soak 4

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