Team:Bielefeld-Germany/Labjournal/Molecular

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<h1>PCR Protocols and Programs</h1>
<h1>PCR Protocols and Programs</h1>
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<h3><a name="cyt_frag1">Phusion 50ul for mtrCAB Fragment1</h3>
<h3><a name="cyt_frag1">Phusion 50ul for mtrCAB Fragment1</h3>

Revision as of 15:20, 31 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 °C98 °C68 °C72 °C72 °C
30s 10s30s 1:356:00
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 °C98 °C60 °C72 °C72 °C
1:00 10s30s 30s6:00
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 °C98 °C70 °C72 °C72 °C
30s 10s30s 50s6:00
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 °C98 °C70 °C72 °C72 °C
45s 30s1:00 45s5:00
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 °C98 °C60 °C72 °C72 °C
30s 10s30s 1:156:15
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 °C98 °C68 °C72 °C72 °C
30s 10s30s 1:356:00
35 cycles
Notes:
Size: 6311bp
Primer1:
Primer2:













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