Transcriptomics

Question 1

Define transcriptomics.

Answer 1

Global analysis of gene expression (RNA).

Question 2

What are the 2 sections of the transcriptomics process?

Answer 2

Sample preparation and data processing.

Question 3

What are the 6 steps of the transcriptomics process?

Answer 3

1) Extract RNA (avoiding degradation).
2) Generate a library.
3) Sequence or perform a microarray.
4) Data processing.
5) Statistical analysis.
6) Functional interpretation.

Question 4

What are confounding factors?

Answer 4

Differences that may affect the results of the study.

Question 5

What must be considered to rule out confounding factors?

Answer 5

Sampling time eg circadian influence.
Operator eg animal handler.
Underlying physiology eg sex, genetic similarity.
Experimental setup eg position in incubator.

Question 6

What is sample heterogeneity and why should it be avoided?

Answer 6

Sample heterogeneity is when different cell types from the same organism are present in a sample. It should be avoided as it makes comparisons difficult.

Question 7

What method is used to prevent sample heterogeneity?

Answer 7

Drop-sequencing. Each individual cell is encapsulated and analysed individually.

Question 8

How can RNA purity be tested?

Answer 8

RNA absorbs light at the 260nm wavelength. Anything absorbing light outside of 250-270nm range isn’t RNA.