12. For each of the following, choose one of the listed experimental approaches

Question

12. For each of the following, choose one of the listed experimental approaches that you think is the most appropriate to answer the question. Possible approaches: ChIP-PCR, Reverse Transcription-qPCR, Western blot, yeast two hybrid, DNase I footprinting, EMSA, immunohistochemistry (8 pt)

a) Is Gene Z transcribed, ie, is the RNA present, in cell type A?

b) Does transcription factor TF bind to a region within 500 base pairs of Gene Z in vivo in cell type A?

c) Does transcription factor TF bind to RNA polymerase?

d) Is transcription factor TF localized in the nucleus?

Explanation / Answer

a) If gene Z is transcribed then Z protein will form in cell type A and western blot technique is most appropriate to check this as it detects the expression of m-rna in form of protein within a cell.

b) To locate exact location or sequence where protein binds within a DNA possible approach is DNase I foot printing assay. If genomic binding site is known it can be checked by ChIP-PCR.

c) Reverse transcription- qPCR is most appropriate way to check either transcription factor bind to RNA polymerase or not.

d) By Yeast two hybrid system we can confirm either transcription factor is localized in nucleus or not.

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