Exam II review sheet 10.31.09

Exam II review sheet 10.31.09 - MCB110 Fall 2009 Exam II...

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MCB110 Fall 2009 Exam II Review Questions For the following experimental techniques, briefly describe the procedure, and exactly what you measure with it (i.e., what does/do the gel/fluorescent signal/bands/etc. tell you) or what is accomplished: Northern Southern DNaseI footprinting assay Co-transfection with a reporter gene EMSA/gel shift Ion-exchange chromatography ChIP Experimental questions: Give at least two ways to determine what polymerase is transcribing a given region of DNA. 1. You are studying a transcription factor MCB1, and you have narrowed its binding site to a 400 base pair sequence using genetic mapping. Unfortunately, your lab has run out of DNase, and so you are unable to perform a footprinting assay. How else could you narrow down the sequence specificity of MCB1? Explain your experiment using diagrams as needed. 2. You successfully identify a 25 base pair binding site for MCB1. What experiment could you do to determine whether this 25 base pair sequence is necessary and sufficient for transcriptional activation by MCB1? (Both in vitro and in vivo experiments are acceptable). 3. You are now curious about the distance of this 25bp element from a transcriptional start site. You know the coding sequence for Mcb2, a gene that is transcriptionally activated by MCB1. Describe one
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This note was uploaded on 02/04/2010 for the course CHEM 113 taught by Professor Toste during the Fall '07 term at Berkeley.

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Exam II review sheet 10.31.09 - MCB110 Fall 2009 Exam II...

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