Preparation of pMPMT5-AID+AIDT7 construct

1. Transformation of E. coli TOP10 strain with ligation from previous day
2. Plating transformants on LB + tetracycline

Polymerase Chain Ligation (PCL)

Product	Templates	Primers	Product length
Alpha-A fusion	Alpha-linker + linker-A	AlphaL+SacI + AP+NotI	1500 bp
Omega-A fusion	Omega-linker + linker-A	OmegaL+SacI + AP+NotI	1440 bp

PCL program for fusion A with alpha and omega

Temperature	Time
94°C	4:00
94°C	0:30	28 cycles
48°C-62°C	0:45
72°C	2:30
72°C	10:00
4°C	infinite