assignment

DNA manipulation worksheet

Define/describe its use or purpose:

Electrophoresis

Restriction endonuclease (why restricted?, why endo? Why -nucl-? Why - ase?)

RFLP

Ethidium Bromide

Probe

Complete:

DNA is ___________________ charged and therefore moves toward the _______________ electrode.

__________________ is needed to carry the charge efficiently through the solution.

Why does a gel of RE digested genomic DNA look like a big smear?

Why does a Southern blot made from genomic DNA gel (smear) above show only specific size bands?

Define:

Vector

cDNA

genomic library

How can you tell if your plasmid has an piece of new DNA inserted?

Look at the map of the R plasmid below. If I have cut genomic DNA with EcoRI and SalI, and want to insert genomic pieces into the plasmid, can I? If I can, what antibiotic resistance will be lost by the bacteria carrying the recombined plasmid? How can I detect the proper plasmid?

If I have genomic DNA cut with PstI and PvuI, and want to insert it into the plasmid shown, can I? If I can, what antibiotic resistance will be lost by the bacteria carrying the recombined plasmid? How can I detect the proper plasmid?