Question

5. A qPCR experiment was done in which Saccharomyces cerevisiae cells were collected before and after treatment with the chemical methyl methanesulfonate (MMS). The two genes for which expression was tested were RFA2 and UBC6. UBC6 is a housekeeping gene. Use the qPCR data given below to answer the following questions.

Condition	untreated RFA2	untreated UBC6	MMS treated RFA2	MMS treated UBC6
Ct value	23.77	23.15	25.44	27.78

A) What type of quantitation will you need to use for this data? Explain why.

B) Calculate ΔCt and ΔΔCt values from the data points provided.

C)Based on your calculated values in part (b) did expression of RFA2 increase or decrease? Would you have anticipated this outcome? Why or why not?

4. (0.5 pt) Another untreated sample looking at RFA2 expression (Sample 2) had a Ct value of 27.07 and was known to start with 1000 template strands of RFA2. Knowing this, how many template strands of RFA2 did your initial untreated RFA2 sample (Sample 1) have to start with?

Answer 1