Revision as of 20:39, 30 August 2013

PCR Protocols and Programs

Phusion 50ul for mtrCAB Fragment1

Reagent	1x	8x
Hf-Buffer 5x	10.0	88.0
dNTPs (10mM)	2.0	17.6
Primer1 (10mM)	1.0	8.8
Primer2 (10mM)	1.0	8.8
Template (16.5ng)	3.0	26.4
DMSO	4.0	35.2
Phusion (0.5u)	1.0	8.8
H2O	26.0	228.8

Program Phusion
98 °C	98 °C	68 °C	72 °C	72 °C
30s	10s	30s	1:35	6:00
	35 cycles
Notes: Size: 1800bp Primer1: Primer2:

Phusion 50ul for mtrCAB Fragment2

Reagent	1x	8x
Hf-Buffer 5x	10.0	88.0
dNTPs (10mM)	2.5	22.0
Primer1 (10mM)	1.0	8.8
Primer2 (10mM)	1.0	8.8
Template (5.5ng/ul)	2.0	17.6
DMSO	3.0	26.4
Phusion (0.5u)	1.0	8.8
H2O	29.5	259.6

Program Phusion
98 °C	98 °C	60 °C	72 °C	72 °C
1:00	10s	30s	30s	6:00
	35 cycles
Notes: Size: 330bp Primer1: Primer2:

Phusion 50ul for mtrCAB Fragment3

Reagent	1x	8x
Hf-Buffer 5x	10.0	88.0
dNTPs (10mM)	2.5	22.0
Primer1 (10mM)	1.0	8.8
Primer2 (10mM)	1.0	8.8
Template (5.5ng/ul)	2.0	17.6
DMSO	3.0	26.4
Phusion (0.5u)	1.0	8.8
H2O	29.5	259.6

Program Phusion
98 °C	98 °C	70 °C	72 °C	72 °C
30s	10s	30s	50s	6:00
	35 cycles
Notes: Size: 3000bp Primer1: Primer2:

Phusion 50ul for psB1C3 with Gibson-overlap for mtrCAB

Reagent	1x	8x
GC-Buffer 5x	10.0	88.0
dNTPs (10mM)	2.0	17.6
Primer1 (10mM)	1.25	11.0
Primer2 (10mM)	1.25	11.0
Template (5.5ng/ul)	1.0	8.8
DMSO	1.5	13.2
Phusion (0.5u)	0.5	4.4
H2O	32.5	286.0

Program Phusion
98 °C	98 °C	70 °C	72 °C	72 °C
45s	30s	1:00	45s	5:00
	35 cycles
Notes: Size: 2070bp Primer1: Primer2:

Phusion 50ul mtrCAB

Reagent	1x	8x
GC-Buffer 5x	10.0	88.0
dNTPs (10mM)	2.5	22.0
Primer1 (10mM)	1.0	8.8
Primer2 (10mM)	1.0	8.8
Template (5.5ng/ul)	2.0	17.6
DMSO	3.0	26.4
Phusion (0.5u)	1.0	8.8
H2O	29.5	259.6

Program Phusion
98 °C	98 °C	70 °C	72 °C	72 °C
45s	30s	1:00	45s	5:00
	35 cycles
Notes: Size: 2070bp Primer1: Primer2:

@@ Line 135: / Line 135: @@
-<h3><a name="cyt_frag3">Phusion 50ul for psB1C3 with Gibson-overlap for mtrCAB </h3>
+<h3><a name="cyt_psb1c3_mtr">Phusion 50ul for psB1C3 with Gibson-overlap for mtrCAB </h3>
 <table border=1 style="float:left; padding-left:10px">
@@ Line 174: / Line 174: @@
 <br><br><br><br><br><br><br><br><br><br><br><br><br>
+<h3><a name="cyt_mtrCAB">Phusion 50ul mtrCAB</h3>
+<table border=1 style="float:left; padding-left:10px">
+      <col width="150">
+      <col width="50">
+      <col width="50">
+<tr> <th>Reagent</th>	        <th>1x</th>	<th>8x</th> </tr>
+<tr> <td>GC-Buffer 5x</td>      <td align="right">10.0</td>	<td align="right">88.0</td> </tr>
+<tr> <td>dNTPs (10mM)</td>  	<td align="right">2.5</td>	<td align="right">22.0</td> </tr>
+<tr> <td>Primer1 (10mM)</td>	<td align="right">1.0</td>	<td align="right">8.8</td> </tr>
+<tr> <td>Primer2 (10mM)</td> 	<td align="right">1.0</td>	<td align="right">8.8</td> </tr>
+<tr> <td>Template (5.5ng/ul)</td><td align="right">2.0</td>	<td align="right">17.6</td> </tr>
+<tr> <td>DMSO</td>	        <td align="right">3.0</td>    <td align="right">26.4</td> </tr>
+<tr> <td>Phusion (0.5u)</td>        <td align="right">1.0</td>	<td align="right">8.8</td> </tr>
+<tr> <td>H2O</td>    	       <td align="right">29.5</td>	<td align="right">259.6</td> </tr>
+</table>
+<table border=1 style="float:left; margin-left:20px">
+      <col width="50">
+      <col width="50">
+      <col width="50">
+      <col width="50">
+      <col width="50">
+<tr><th colspan="5">Program Phusion</th></tr>
+<tr align="center"><td>98 °C</td><td>98 °C</td><td>70 °C</td><td>72 °C</td><td>72 °C</td></tr>
+<tr align="center"><td>45s</td> <td>30s</td><td>1:00</td> <td>45s</td><td>5:00</td></tr>
+<tr><td></td><td colspan="3" align="center">35 cycles</td><td></td></tr>
+<tr><td colspan="5">
+<b><u>Notes:</u></b>
+<br>Size: 2070bp
+<br>Primer1:
+<br>Primer2:
+<br>
+</td></tr>
+</table>
+<br><br><br><br><br><br><br><br><br><br><br><br><br>
 </html>

Team:Bielefeld-Germany/Labjournal/Molecular

From 2013.igem.org

Revision as of 20:39, 30 August 2013

PCR Protocols and Programs

Phusion 50ul for mtrCAB Fragment1

Phusion 50ul for mtrCAB Fragment2

Phusion 50ul for mtrCAB Fragment3

Phusion 50ul for psB1C3 with Gibson-overlap for mtrCAB

Phusion 50ul mtrCAB