Reporter + Blot Analysis

Question 1

How do you use reporter analysis to measure promotion

Answer 1

Fuse a promoter with a reporter gene and can detect levels of expression/promotion

Question 2

How do you work out which part of a sequence is important for promotion using reporter analysis

Answer 2

You can truncate the promoter region via deletion and see changes in levels of expression. If truncation affects it that’s where the promoter sequence is

Question 3

What is northern blot used for

Answer 3

Measuring rna levels

Question 4

What happens to the rna in northern blotting

Answer 4

Separated in gel electrophoresis then the bands are transferred to a membrane and lighted with a probe

If bands are darker it means more mrna/rna

Question 5

What is needed to measure levels of mrna in northern blot

Answer 5

Compare to a loading control/reference gene which has constant levels of expression

Question 6

What is western blot used to measure

Answer 6

Levels of protein

Question 7

What happens in western blotting

Answer 7

Transfer a protein to a membrane and denature via page

Protein attaches to a primary antibody recognised by a secondary antibody with enzyme

Substrate added and shows protein levels

Question 8

Like northern blotting, what is needed as a control to detect levels of protein in western blot

Answer 8

Loading control/reference gene where protein is produced at constitutive levels