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From 2013.igem.org

Contents 1 208 1.1 Main purpose 1.2 Who was in the lab 1.3 Procedure 1.3.1 Combine Nir operon with pZA21

208

Main purpose

• PCR with USER primers to combine Nir operon with pZA21

Who was in the lab

Ariadni,Henrike,Julia,Jakob

Procedure

Combine Nir operon with pZA21

Following Mathildes Protocol for DpnI digestion of our linear pZA21 w. Uracil incerts

Mastermix for 7 PCR tubes, Using the mastermix protocol we got from Mathilde.

triplets of each of the two Nir Operon extracts that we have made yesterday, including one negative control:

• Nir 2a(I,II,III)
• Nir 2b(I,II,III)

Using the following PCR program:

	time	Temp C
Denaturing	2 min	98
start of loop	10 sec	98
annealing	30 sec	60
elongation	9 min	72
goto start of loop 35 times
hold		10