For this project you will need to find out as much as possible about a plasmid that will be given to your group. All that is known about the plasmid is that it can replicate in E. coli, it carries the ampicillin resistance gene and has a single Eco RI site.
Initially, you will need to decide which questions need to be answered about
the plasmid. Write all of them down and then decide which will be easiest to
find the answers to first. You will probably not have time to answer all the
questions so you need to consider those questions that will give you the most
information about this plasmid per time invested.
You will be given 50 ng of purified plasmid DNA. From this DNA you will need
to develop an experimental design to find out as much about the plasmid as
possible in the five weeks that you will be given for this project.
The techniques that you have available to you include standard procedures
for DNA amplification, purification and quantification. Analytical techniques
include various Restriction Enzyme Analysis, DNA sequencing, Sub-cloning,
Primer design, PCR, and Computer Sequence Comparison Programs.