Molecular Biology-2014
Assignment #2
Exercise 1
Dilutions:
1. Indicate how you prepared 1 mL of each of the following solutions:
A 1.5mM solution of compound “A”.
A 0.36% (m/v) solution of compound “B”.
A 6% (v/v) solution of solution I.
A solution containing 0.5mg of compound “A” and 0.1% (v/v) of
compound “B”.
e. A solution representing the following ratio: solution I : solution II : water :
2 :1 :2
a.
b.
c.
d.
You do not need to show your calculations. Simply indicate the volume of each
component added.
2. Indicate the average absorbencies of each of the solutions you prepared.
Bradford assay:
3. Submit a standard curve of absorbance Vs. the standard concentrations of BSA in
mg/mL. The graph should be generated with excel. Both axes should be linear scales.
4. Based on the absorbance readings obtained for the BSA solution of unknown
concentration, what was the concentration of the unknown BSA solution?
Agarose gel electrophoresis:
5. Submit a figure representing your agarose gel. Make sure to include an appropriate
legend. Follow the directives for figures on the web page of this course. Make sure to
include all the required information in the legend for the understanding and
interpretation of the figure.
6. Submit a standard curve of the molecular weight ladder. Follow the directives for
generating such a curve under the heading Graphs on this course's web site.
7. Submit a table presenting the analysis of the restriction digests. Your table should
include: Enzyme used, Total number of cuts, Number of cuts in the vector, Number
of cuts in the insert, and Fragments sizes generated.
8. In the figure legend to the figure of your gel provide a brief analysis indicating the
following information: Total plasmid size, Size of the insert and the restriction site
representing the insertion site.
9. Submit a figure of the restriction map of the insert. Your map must be linear, include
the multiple cloning site, indicate the insertion site, include the positions in the
multiple cloning site or the insert of all the enzymes tested. Your figure must be to
scale. Follow the directives for generating such a figure under the heading
Graphs/Figures on this course's web site.
Molecular Biology-2014
Exercise 2
Plasmid DNA isolation and agarose gel electrophoresis:
1. Submit a figure representing your agarose gel. Make sure to include an appropriate
legend. Follow the directives for figures on the web page of this course. Make sure to
include all the required information in the legend for the understanding and
interpretation of the figure.
2. Submit a table indicating the absorbencies at 260nm of your dilutions of salmon
sperm DNA and the corresponding concentrations in µg/mL.
Plasmid DNA digestion:
3. Based on the results obtained from your restriction digests, answer the following
questions:
a. How many times did PvuII cut within the plasmid?
b. How many times did HincII cut within the plasmid?
c. How many times did HincII cut within the PvuII fragment?
d. What are the distances between the PvuII and the HincII sites?
Bioinformatics 1:
5. Submit a table with the following information with regards to each of the unknown
genes from the first bioinformatics exercise.








The accession number (#2)
Coverage
Max Ident.
E value
The definition (#1)
The organism from which this sequence was obtained (#3)
The product of the gene (#4)
The protein id. This is the protein’s accession number (#5)
6. Submit a printout of a FASTA sequence of one of the unknown genes. Include the
name of the gene as a heading to the printout.