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The true test of a first-rate mind is the ability to hold
two contradictory ideas at the same time.
F. Scott Fitzgerald

Day 5: Construction of a simple circuit

Assignments Due

Overview of Experiment

Today you will begin building your new circuit. Lab sessions 5-8 are self-scheduled (lab "begins"after the presentations and "ends" at 6 p.m.) ; if you need to come in at other times (9:30 a.m. - 5:30 p.m., weekdays only!) / days to pull plates or start overnight cultures, please let me know so I can unlock the labs.

Before beginning procedures on Day 5, record an outline of your planned experimental strategy for building the simple circuit (described below) in your lab notebook. Include details such as restriction enzymes you will use and how you will confirm the identity of your construct (Hint: we identified transformants TWO different ways in lab) as well as expected results.


Plasmid DNA mini preps and restriction enzyme digests are highly recommended to screen putative transformants: pick isolated colonies and grow overnight in 4 ml LB + Amp; use 3 ml for mini prep and make glycerol stock of 1 ml. If you decide to screen by colony PCR, you do not need a (+) control for PCR; however, you should run a (-) control reaction. 


Copyright, Acknowledgements, and Intended Use
Created by B. Beason (bbeason@rice.edu), Rice University, 21 November 2007
Updated 4 November 2009