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.)

Chromosome Translocation: chromosome microarray + FISH? + CGH?
Determination of the levels of gene expression of Gene XYZ with or without hormone treatment in a cell line: FISH + (RT-PCR)
Presence of a K-ras mutation in a set of head and neck tumors: Sanger + Histology + NGS?
Gene Amplification: PCR + FISH? + qPCR
Identifying CD4 and CD8 positive T cells: Flow cytometry + immunohistochemistry

Can someone please verify my answers? Thanks.

Explanation / Answer

Chromosome translocation: Chromosome microarray can detect microdeletion, microinsertion and unbalanced translocation. It cannot detect balanced translocations and chromosomal inversion. Fluoresence in situ hybridisation (FISH) can detect both constitutional and acquired chromosomal aberrations. FISH can be applied either to metaphase or interphase chromosome. Unknown familial genetic polymorphisms may result in false positive or negative FISH results. Comparative genomic hybridization (CGH) testing is useful for the identification of specific genes involved and in sizing a chromosomal abnormality detected by conventional cytogenetics or FISH techniques.

Gene expression: RNA FISH can be used to study gene expression. The many advantages of using FISH in studying gene expression are absolute quantification, high time resolution, use of wild-type cells, ability to simultaneously measure multiple mRNA types, and broad application across species. Reverse transcription PCR (RT-PCR) is considered as the gold standard for accurate, sensitive and fast measurement of gene expression without hormone treatment in a cell line.

Detection of k-ras mutation: The targeted next generation sequencing (NGS) platform is specific and sensitive for KRAS exon 2 mutation detection and is appropriate for use in routine clinical testing. It is cost-efficient and time-efficient. Sanger sequencing for detecting k-ras mutation is time consuming and non specific. Immunohistochemistry and histopathological diagnosis are used as screening tests.

Gene amplification: Polymerase Chain Reaction (PCR) amplifies single copy or several copies of segment of DNA. PCR monoplexing appears to have the advantage of more accurately quantitating high levels. Gene amplification by FISH analysis has the advantage of allowing concurrent assessment and architecture.

CD4 and CD8 T cells: Flow cytometry is a technique to count and sort cells. It is simple, rapid and easy to perform. Density and distribution of CD4 and CD8 cells can be found using immunohistochemistry.

Related Questions

Navigate

• Browse (All)
• Browse G
• Subjects

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.