PCR Screening of L173 transformants

Transformation results (rbs-TetR in mRFP-LVA-tripart-pSB1A3)

Ligation L173

• insert: D112 (ES) rbs-TetR
• vector: D187 (EX) mRFP-LVA-tripart-pSB1A3

PCR Screening

PCR screening

PCR screening programm

• elongation tim: 2 min
• primers used: O18 & O19
• positive PCR control: S158 (pSB3K3)
• negative PCR control: no template

Results: the clones are not correct.

Digestion

• 5 µL of DNA
• 3 µL of 10X buffer n°2
• 0,3 µL 100X BSA
• 1 µL of EcoRI
• 1 µL of SpeI
• 19,7 µL of water

Incubation 2h30 at 37°C and then 20 min at 65°C.
Not purified yet. Put at -20°C.

Transformation results

L177 = d134 + d187 (pFlhB-mRFP LVA+)

L175 = d149 + d° (pFliL - ECFP LVA+)

L176 = d149 + d° (pFliL - ECFP LVA-)