5/17

Transform

LB plate (Ampicillin resistance)

RhlR (Bba_C0171),

pRhlR (Bba_R0071),

LasR (Bba_C0179),

pLas (Bba_R0079),

mCherry (Bba_J06504),

mRFP (Bba_E1010),

GFPmut3 (Bba_E0040),

Pc (Bba_J23119),

double terminator (B0015),

RBS (B0030)

5/18

Inoculation & Incubation：LB broth(Amp)

RhlR (Bba_C0171),

pRhlR (Bba_R0071),

LasR (Bba_C0179),

pLas (Bba_R0079),

mCherry (Bba_J06504),

mRFP (Bba_E1010),

GFPmut3 (Bba_E0040),

Pc (Bba_J23119),

double terminator (B0015),

RBS (B0030)

5/19

Get Plasmid: Use Plasmid miniprep purification kit

RhlR (Bba_C0171),

pRhlR (Bba_R0071),

LasR (Bba_C0179),

pLas (Bba_R0079),

mCherry (Bba_J06504),

mRFP (Bba_E1010),

GFPmut3 (Bba_E0040),

Pc (Bba_J23119),

double terminator (B0015),

RBS (B0030)

5/22

Digestion:

B0030

mCherry

mGFP

RFP

LasR

RhlR

After digestion we operate gel electrophoresis:

Ligation overnight

B0030v-mCherry

B0030v-mGFP

B0030v-RFP

B0030 v-LasR

B0030v-RhlR

5/23

Transform:

B0030-mCherry

B0030-mGFP

B0030-RFP

B0030-LasR

B0030-RhlR

5/27

Inoculation & Incubation：

LB broth(Amp), 5 tubes for each kind

B0030-mCherry

B0030-mGFP

B0030-RFP

B0030-LasR

B0030-RhlR

5/28

Check:

B0030-mCherry

B0030-mGFP

B0030-RFP

B0030-LasR

B0030-RhlR

5/30

Get Plasmid:

Use Plasmid miniprep purification kit

B0030-mCherry

B0030-mGFP

B0030-RFP

B0030-LasR

B0030-RhlR