Download S3. pClone Intro-Weekly Questions for Students to

Using Synthetic Biology and pClone Red for Authentic Research on Promoter
Function: Introductory Biology (identifying new promoters)
A. Malcolm Campbell and Todd T. Eckdahl
Weekly Questions For Students to Answer Prior to Lab
Week 1
Answer each of these four questions in three sentences or less.
A) What does the promoter of a gene do?
B) What is a plasmid?
C) What are oligonucleotides (often referred to as oligos)?
D) What is a restriction enzyme?
Week 2
Answer each of these four questions in three sentences or less.
A) How are type IIs restriction enzymes different from the more commonly used type II
restriction enzymes?
B) Is Bsa I a type II or type IIs restriction enzyme? How did you find out?
C) What is DNA ligase?
D) What is bacterial transformation with regards to DNA engineering?
Week 3
Answer each of these four questions in three sentences or less.
A) How will you ligate your new promoter into a plasmid for testing?
B) What will the plasmid need to contain if you want to determine if your promoter is
working?
C) How is fluorescence of red fluorescent protein (RFP) measured?
D) How is a spectrophotometer used to measure cell density in a population of E. coli?
Week 4
Answer each of these three questions in three sentences or less.
A) Are promoters more like on/off switches, or rheostats?
B) What could you do to be sure you successfully cloned the promoter you wanted to
clone?
C) How will you calculate the level of fluorescence of your cells to take into account that
different treatments may produce different densities of cells?
Week 5
Answer each of these three questions in three sentences or less.
A) If your promoter did not work the way you hypothesized, what should you do for your
presentation?
B) What did you learn about your promoter that you built from scratch?
C) What sort of information is encoded in a promoter sequence?