L21 Flashcards
(21 cards)
EtBr glows when
it is in between the DNA base pairs
largest bp (agarose gel) that can run
20 000 bp
star activity
reduced specificity in recognizing base pairs (wrong locations will be cut)
partial digest
pieces we want, and combinations that are still stuck together (bands will show as larger than they should)
star activity is more cuts or less
more because they cut what they should and more
difference between incomplete digest and star activity on a gel
incomplete is bigger bands that are fainter
star activity smaller bands that are fainter
pulsed field gel electrophoresis
alternates the direction in order to separate large DNA pieces (up to 1Mb)
the _______ the DNA, the easier to break
longer
shearing
just snapping DNA into smaller pieces
PulseNet
created to do surveillance/tracking of foodborne illnesses
how are foodborne illnesses studied
PFGE (pulse field gel electrophoresis)
what enzyme connects phosphodiester backbone of DNA between the 3’OH and 5’ P
ligase
what does ligase need to stick together
3’OH and 5’ P
how might plasmids enter cells
heat shock to invert membrane and uptake plasmids
do cells need recovery time after plasmids enter
yes
the “loading” sport is typically a
multiple cloning site
where is the multiple cloning site located
within insert differation gene
X gal colonies will turn ____ when its an empty vector and the enzyme is made
blue
does a white air blue X gal indicate success? why?
white indicates success because the insert is present (lacZ gene is disrupted)
what is plasmid extraction called
miniprep because you only make a few ml with these colonies
does plasmid DNA stay in supernatant
yes
