NYMU Taipei/Lab Notes/2007 9 30

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< NYMU Taipei/Lab Notes(Difference between revisions)
 
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<p>
 
<h4>PCR for TAT_A-INS_A TAT_B-INS_B</h4>
<h4>PCR for TAT_A-INS_A TAT_B-INS_B</h4>
{| align=left | border=1 | style="text-align:center"
{| align=left | border=1 | style="text-align:center"
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|1
|1
|TAT_A-INS_A
|TAT_A-INS_A
-
|
+
|209
|TAT_F+INS_A_R
|TAT_F+INS_A_R
|TAT_A,INS_A PCR product@9/29
|TAT_A,INS_A PCR product@9/29
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|2
|2
|TAT_B-INS_B
|TAT_B-INS_B
-
|
+
|233
|TAT_F+INS_B_R
|TAT_F+INS_B_R
|TAT_B,INS_B PCR product@9/29
|TAT_B,INS_B PCR product@9/29
|-
|-
|}
|}
-
</p>
+
 
 +
{|align=left| border=1 | style="text-align:center"
 +
 
 +
|Template1
 +
|1uL
 +
|-
 +
|Template2
 +
|1uL
 +
|-
 +
|10X buffer
 +
|5uL
 +
|-
 +
|2.5mM dNTP
 +
|2uL
 +
|-
 +
|Primer_F
 +
|1.5uL
 +
|-
 +
|Primer_R
 +
|1.5uL
 +
|-
 +
|<i>Pfu</i> polymerase
 +
|0.5uL
 +
|-
 +
|ddH2O
 +
|37.5uL
 +
|-
 +
|Total
 +
|50uL
 +
|-}
 +
 
 +
 
 +
{| align=left | border=1 | style="text-align:center"
 +
!Step
 +
!Temp (°C)
 +
!Time (mins)
 +
!Cycle
 +
|-
 +
|Initial Denaturation
 +
|95
 +
|10
 +
|
 +
|-
 +
 
 +
|Denatuation<br>Annealing<br>Extension
 +
|95<br>55<br>72
 +
|1<br>1<br>1
 +
|35
 +
|-
 +
 
 +
 
 +
|Final Extension
 +
|72
 +
|10
 +
|
 +
|-
 +
 
 +
|Soak
 +
|4
 +
|
 +
|
 +
|-
 +
|}
 +
 
 +
 
 +
 
 +
<br><br>
<h4>2%Gel check for TAT_A-INS_A TAT_B-INS_B</h4>
<h4>2%Gel check for TAT_A-INS_A TAT_B-INS_B</h4>

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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