Research Methodology & Design Flashcards

Which guideline sets out the minimum information that should be reported for qPCR experiments in publications?

Answer: MIQE

MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) defines qPCR reporting standards.

A researcher runs each cDNA sample in triplicate wells on one plate. What do these replicates mainly measure?

Answer: Pipetting and instrument precision

Technical replicates from the same sample capture assay precision, not biological variability.

What is the main purpose of a no-reverse-transcriptase (no-RT) control in an RT-qPCR study?

Answer: To detect genomic DNA contamination in RNA samples

Amplification in a no-RT control means DNA, not cDNA, served as template.

For an expression study comparing treated and untreated mice, which unit should be used as the experimental unit for statistics?

Answer: Each individual mouse

Independent biological subjects are the experimental units; wells are not independent observations.

Placing samples from all treatment groups on every plate rather than one group per plate is an example of what design principle?

Answer: Blocking

Blocking by plate prevents plate-to-plate variation from being confounded with treatment.

Before choosing reference genes for a new tissue and treatment, what is the best practice?

Answer: Validate candidate genes for stability under the study conditions

Reference gene stability varies by tissue and treatment and must be empirically validated, e.g., with geNorm or NormFinder.

A power analysis performed before a PCR study is primarily used to determine what?

Answer: The number of biological samples needed to detect an expected effect

Power analysis estimates sample size needed to detect a specified effect with acceptable error rates.