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Lab 208

Main purpose

New PCR on PAO1 for extraction of Nir - again again. USER reaction between pZA21::RFP no promoter USER fragment and the araBAD USER fragment PCR of pZE21 to switch out the native promoter with araBAD PCR on HAO

Who was in the lab

Kristian

Procedure

PCR

• pZE21 no promoter with primer pair 13, 57C and 3:00
• HAO with primer pair 18, 59C and 3:00
• Nir by colony PCR on the 4 new plates with PAO1. Primer pair 41 and 42 where used, both on touchdown PCR 71→61 1C increment 10 cycles and then additional 25 cycles on 61C.

USER-reaction

With pZA21::RFP no promoter and the araBAD USER fragment plus two controls; one with araBAD fragment only and one with pZA21::RFP fragment only.

Results

See gels on tomorrow's page.

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