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60 Cards in this Set

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  • Back
_______,the cellular absorption and expression of the new gene combination in a cell
transformation
Because the genetic code is nearly universal,________can accept foreign DNA derived from any other organism.
e.coli
A foreign gene inside e.coli can be replicated, and in most cases translated, into a __________
protein
e.coli is a normal inhabitant of the human__________
digestive tract
in the large intestines, e.coli absorbs digested foods and produces______that are useful to humans.
vitamins
it makes sense to incubate e.coli cells at the body temperature of___________
37
__________contains nutrients similar to those found in the colon.
luria burtoni broth
the natural transformation of bacterial cells, was first described by____________, in 1928.
frederick griffith
e.coli cells growing in mid-log growth phase, in a cold________ solution are heat shocked at 42c
calcium chloride
some bacteria that are actively dividing contain membrane___________that allow the passage of DNA
PORES
positively charged calcium ions seem to bind_________in the cell membrane at the location of the pore.
neg, phospholipids
calcium ions seem to attract negatively charged_________to that region of the cell.
DNA
_______may physically pump DNA through the pore.
heat shocking
_____________can destroy the antibiotic, ampicillin.
penicillinase
x-gal is a substrate analog of lactose that forms a______________ color when digested.
blue
e.coli cells that have been transformed with___________ will form blue colonies when grown on media containing x-gal.
pBLU
in the absence of________x-gal will not be digested and bacterial colonies will remain white.
`beta galactosidase
white colonies that grow on media containing ampicillin and x-gal are those that have absorbed the________complex
r-p VIB
the ability of transformed e.coli cells to grow on media containing ampicillin is one of two markers used to ________transformed cells.
select
what is the first reagent added to the transformation tubes?
calcium chloride
what is added to the tubes after the heat shock treatment?
luria burtani broth
growth of colonies on antibiotic medium provides______ evidence that bacterial cells have been transformed with a plasmid.
phenotypic
to confirm this at the _________level, plasmid DNA is isolated from the transformants.
genotypic
restriction analysis of recombinant DNA provides a_____ of its genetic identity.
fingerprint
transformed cells taken from an antibiotic resistant colony are grown to _________phase in a overnight culture,
stationary
cell are collected by_______
centrifugation
ethylenediamine tetra acetic acid (EDTA) binds positive calcium and magnesium ions that are necessary for___________stability.
membrane
_________, an ionic detergent, dissolves the phospholipid and protein components of the cell membrane.
SDS
______________denatures both plasmid and chromosomal DNAs into single strands.
sodium hydroxide
the single stranded__________loops remain linked together like interlocking rings.
plasmid
at a neutral pH, the DNAs______________
recombine
____________DNA only partially recombine and becomes trapped in the SDS/lipid/protein precipitate.
chromosomal
the plasmid DNA recomnbines into double stranded molecules that remain in______________
solution
_____________is added to precipitate the plasmid DNA, which is pelleted by centrifugation.
isopropanal
the pellet containing the plasmid is washed with____________and resuspended in a small volume of TE buffers/
ethanol
treatment with__________destroys RNA, leaving clean plasmid DNA
RNASE
What remains in the tube, as a pellet, after step#4?
cells
what remains in the tube as a pellet after step #14
DNA
how long do the cells incubate in isoprpanol at room temp?_____________
2 min
what is used to dry the nucleic acid pellet?>
kair dryer
in what buffer is the DNA precipitate stored>?>
T.E
Dna molecules of different___________can be separated using the process of gel electrophoresis.
different sizes
the name for this technique means that DNA molecules will be carried with__________through a jell like material.
electricity
DNA molecules are nucleic acids with a___________electrical charge.
neg
when placed in an electrical field, DNA molecules will move towards the ___________pole.
pos
during the electrophoresis process, DNA fragments sort by size in____________
agarose gel
the porous gel matrix acts as a__________
molecular sieve
the distance traveled by a DNA fragment is____________proportional to its size.
inversely
in a given period of time smaller DNA fragments travel___________through the gel.
further
a gel can efficiently separate DNA fragments ranging in size from 100 nuclotides to more than__________
50,000
DNA fragments of different size ranges can be seperated by adjusting the____________of agarose.
concentration
a low concentration (down to 0.3%) produces a loose gel that is more appropriate for separating___________fragments.
larger
a dense sucrose solution causes the DNA to_______into a depression in the gel
sink
_______________in the loading solution, which has a negative electrical charge, move through the ge; with the unseen DNA
visible dyes
following electrophoresis, the gel is soaked in a solution of the stain,_____________
ethidium bromide
the stain becomes concentrated in regions where_______exist.
DNA fragments
the stained gel is exposed to__________
uv light
stained DNA fragments appear as_________ fluorescent bands in the gel.
orange
each DNA band is a collection of millions of identical DNA molecules, all of the same_________
nucleotide length
at what voltage will the power source be set?
150 volts