Biotech lab Q's? ASAP.?

Question

1. Some of the circular lambda molecules are colvalently linked at the COS sites. Other circlets are only hydrogen-vonded and can dissociate to form linear molecules. Heating Lambda DNA to 65 degree C for 10 minutes linearizes any noncovalent COS circlets in the preparation by breaking the hydrogen bonds that hold the complementary COS sites together.

a. Set up duplicate restriction digests of lambda DNA with serveral enzymes. Then heat one reaction from each set at 65 C for 10 mins. while holding the duplicates on ice. Relate changes in restriction patterns of heated versus unheated DNA to a restriction map of the circular lambda genome.
b. how can the data generated by this experiment be used to quantify the appros. % of circular DNA in your preparation?

2. Design and carry out a series of experiments to study the kinetics of a restriction reaction.
a. Determine approx. % of digested DNA at various time points
b. repeat experiments with serveral enzyme and DNA dilutions.

2c. In each case, at what time point does the reaction appear to be complete?

3. Design and test an assay to determine the relative stabiloty of BamHI, EcoRI, and HindIII at room temperature.

4. Determine the identity of an unknown restriction enzyme.
a. Perform single digests of Lambda DNA with the unknown enzyme, as well as with serveral known restrictoin enzymes. Run teh restiction fragmenst in an agarose gel at 50 volts to produce well-spread and well-focused bands.
b. for each fragment, plot dstance migrated vs. base-pair size.

5. research the steps needed to purify a restriction enzyme from E.Coli and characterize its recogniction sequence.



ANYONE WITH AN ANSWER TO ANY ONE OF THESE QUESTIONS IS APPREICATED!

Answer 1

Sorry but you'll have to do your own homework