Given the following scenarios name 2 methods that could be used to investigate t

Question

Given the following scenarios name 2 methods that could be used to investigate these experimental situations and name the strengths and weaknesses of each. (Note there may be more than 2 techniques relevant to the scenario proposed.)

1) Chromosome Translocation: chromosome microarray + FISH? + CGH?

2) Determination of the levels of gene expression of Gene XYZ with or without hormone treatment in a cell line: FISH + (RT-PCR)

3) Presence of a K-ras mutation in a set of head and neck tumors: Sanger + Histology + NGS?

4) Gene Amplification: PCR + FISH? + qPCR Identifying CD4 and CD8 positive T cells: Flow cytometry + immunohistochemistry

My answers are in bold, however, I'm not positive they're correct. If someone can please look at my answers and either confirm the accuracy or correct them if wrong. Thanks so much in advance.

Explanation / Answer

1) Chromosome Translocation: chromosome microarray + FISH? + CGH

Correct.

2) Determination of the levels of gene expression of Gene XYZ with or without hormone treatment in a cell line: FISH + (RT-PCR)

this would better be done by RT- qPCR, where reverse transcription is done and is followed by quantitative PCR. The quantification of gene level expression can also be done by protein quantification profiling with horseradish peroxidase or antibody conjugate or RNA fish.

3) Presence of a K-ras mutation in a set of head and neck tumors: Sanger + Histology + NGS?

Methods to be used are Direct sequencing or Sanger sequencing, the amplification refractory mutation system using a bifunctional self-probing primer (ARMS/Scorpion, ARMS/S) and high resolution melting technique.

4) Gene Amplification: PCR + FISH? + qPCR Identifying CD4 and CD8 positive T cells: Flow cytometry + immunohistochemistry

correct .

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