Team:Bielefeld-Germany/Labjournal/Molecular

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Genetic enginering protocols

Standard Protocols

[Standard Protocols] Standard protocols

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

Specific Protocols

[Cytochromes] 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:

[Cytochromes] 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:

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