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	<script>		<script>
	$(document).ready(function(){		$(document).ready(function(){
-	$(.cyt_frag1_headline).click(function(){	+	$("#item").click(function(){
	$(.cyt_frag1).toggle();		$(.cyt_frag1).toggle();
	});		});

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Revision as of 15:09, 31 August 2013

PCR Protocols and Programs

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 and without 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_Gibson: Primer2_Gibson: Primer1: Primer2:

Phusion 50ul for mtrCAB cluster

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
30s	10s	30s	1:15	6:15
	35 cycles
Notes: Size: 5150bp Primer1: Primer2:

Phusion 50ul for ccmAH cluster

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 (5.5ng/ul)	3.0	26.4
DMSO	3.0	26.4
Phusion (0.5u)	1.0	8.8
H2O	29.5	255.2

Program Phusion
98 °C	98 °C	68 °C	72 °C	72 °C
30s	10s	30s	1:35	6:00
	35 cycles
Notes: Size: 6311bp Primer1: Primer2: