NYMU Taipei/Lab Notes/2007 9 30

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< NYMU Taipei/Lab Notes(Difference between revisions)
 
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<h4>2%Gel check for TAT_A-INS_A</h4>
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<h4>PCR for TAT_A-INS_A TAT_B-INS_B</h4>
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[Image:NYMU_Taipei_20070930_TATA-INSA_check.jpg]
+
{| align=left | border=1 | style="text-align:center"
 +
!
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!
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!size
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!Primer
 +
!Template
 +
|-
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|1
 +
|TAT_A-INS_A
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|209
 +
|TAT_F+INS_A_R
 +
|TAT_A,INS_A PCR product@9/29
 +
|-
 +
 
 +
|2
 +
|TAT_B-INS_B
 +
|233
 +
|TAT_F+INS_B_R
 +
|TAT_B,INS_B PCR product@9/29
 +
|-
 +
|}
 +
 
 +
{|align=left| border=1 | style="text-align:center"
 +
 
 +
|Template1
 +
|1uL
 +
|-
 +
|Template2
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|1uL
 +
|-
 +
|10X buffer
 +
|5uL
 +
|-
 +
|2.5mM dNTP
 +
|2uL
 +
|-
 +
|Primer_F
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|1.5uL
 +
|-
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|Primer_R
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|1.5uL
 +
|-
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|<i>Pfu</i> polymerase
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|0.5uL
 +
|-
 +
|ddH2O
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|37.5uL
 +
|-
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|Total
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|50uL
 +
|-}
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 +
 
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{| align=left | 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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|Initial Denaturation
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|95
 +
|10
 +
|
 +
|-
 +
 
 +
|Denatuation<br>Annealing<br>Extension
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|95<br>55<br>72
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|1<br>1<br>1
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|35
 +
|-
 +
 
 +
 
 +
|Final Extension
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|72
 +
|10
 +
|
 +
|-
 +
 
 +
|Soak
 +
|4
 +
|
 +
|
 +
|-
 +
|}
 +
 
 +
 
 +
 
 +
<br><br>
 +
 
 +
<h4>2%Gel check for TAT_A-INS_A TAT_B-INS_B</h4>
 +
[[Image:NYMU_Taipei_20070930_TATA-INSA_check.jpg|300px]]
 +
*Well 1 TAT_A-INS_A  10uL
 +
*Well 2 TAT_B-INS_B  10uL
 +
*Well 3 100bp ladder  5uL

Latest revision as of 13:29, 1 October 2007

PCR for TAT_A-INS_A TAT_B-INS_B

size Primer Template
1 TAT_A-INS_A 209 TAT_F+INS_A_R TAT_A,INS_A PCR product@9/29
2 TAT_B-INS_B 233 TAT_F+INS_B_R TAT_B,INS_B PCR product@9/29
Template1 1uL
Template2 1uL
10X buffer 5uL
2.5mM dNTP 2uL
Primer_F 1.5uL
Primer_R 1.5uL
Pfu polymerase 0.5uL
ddH2O 37.5uL
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




2%Gel check for TAT_A-INS_A TAT_B-INS_B

NYMU Taipei 20070930 TATA-INSA check.jpg

  • Well 1 TAT_A-INS_A 10uL
  • Well 2 TAT_B-INS_B 10uL
  • Well 3 100bp ladder 5uL
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