1 compare samples 13 what is the purpose of including siRNA

1. compare samples 1-3 . what is the purpose of including siRNA oligonucleotides against firefly luciferase? does the level of BTF expression change in the samples? does the level of beta actin change? what can conclude from the first three samples?

2. compare 4-6, what happens to the expression level of BTF in these samples? how does this correlate the siRNA amounts used in these samples? what can you conclude from this experiment with regard to treatment of cells with siRNA oligonucleotides against BTF?

The experimental set up is as follows: Lane 1-Transfection reagent alone these cells Lane 2 6 pmol siRNA\'s against luciferase which does not exist in Lane 3- 60 pmol siRNA\'s against luciferase which does not exist in these cells Lane 4 0.6 pmol Btf siRNA Lane 5- 6 pmol Btf siRNA. Lane 6- 60 pmol Btf siRNA kDa 250 150 100 B-actin 37 1 2 3 4 5 6

Solution

1. the siRNA\'s purpose in any experiment is to interfere with the transcription of the gene it is designed against the gene it is complimentary to, in this case it is the luciferase, it interferes with the gene expression by degrading the mrna of that gene after its transcription

in the given question samples 2 and 3 do not contain the siRNA\'s, and so the expression of BTF and beta actin is intact because the siRNA is not present in the sampples and therefore does not interfere with their expression or degrade their mRNA.