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79 Cards in this Set

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isoelectric focusing is the
1. prots can be sep on basis of their rel contents of acidic and basic residues.

-isoelectric point of a prot is the pH at which the net charge is 0.

-separation of prots in a pH gradient gel

-can resolve prots that differ in pI by as little as 0.01 which means that prots tht differ by one net charge can be separated

2.isolectric point focusing can be combined w/SDS PAGE to obtain high resolution separations
-subject a single sample to isoelectric focusing in a horizontal direction
-single lane gel is placed horizontally on top of SDS PAGE slab and are separated in a vertical direction on the basis of mass

1. strand separat
2. bind of DNA poly that moves in 5-->3
3. strand syn in 5-->3 direction brings nucleotides into make a newly synthesized strand of template
4. Half of the molecule is conserved and rest is newly synthesized
isoelectric point of a prot is the
pH at which the net charge is 0
plasmid purificat
enrich plasmid DNA over chromosomal DNA
purines have
2 rings
sodium dodecyl sulfate, an ionic detergent that binds to prot giving it a negative charge, unfolds 3º sturcture, solubilizes proteins
pyrimidines have
1 ring
DNA->mRNA (nucleus)
2 D gel electrophoresis
prots can be sep on basis of their relative contents of acidic and basic residues


complementary to the template
A260/A280 of pure DNA is between
1.8 and 1.9
prot molecules produce by beta cells of vertebrates(immunoglobulins) and bind to entities called antigens w/high specificity(light and dark chains)
A320>5% of A260, there is a presence of
transmittance and concentration are
inversely proportional
restriction digests
loading dye, plasmids, water, restriction enzymes
nucleotides have
covalent bonds
ciliary motion is due to
bending of ciliary axoneme, as a result of the sliding of the axonemal mts
quaternary structure
2 or more indep folded prot chains loosely held together by weak bonds
plasmid purificat(brief)
phosphate buffer, SDS, potassium acetate, etOH, Tris/EDTA
one helical turn in DNA is
3.4 nm
bacterial transformat
induce competence w/ice cold CaCl2, heat shock high temp for bact envel to become susceptible to uptake of DNA, pUC vector placed in competent E.coli cells: mercaptoetOH to competent cells, add pUC plasmid, heat shock, SOC, plate
G and C have
3 bonds
low ratio of A260/A280
2º antibody recognizes
1º antibody coupled w/dye and is where fluorescence is coming from
DNA extraction
pronase, NaCl, RNase, etOH, Tris/EDTA
a bundle of mts(comprise the cilia and flagella), has 9 pairs of outer mts surr by 2 central mts enclosed by a plasma membrane
tetrahymena are
fresh water protozoan living in axenic culture
last part has + or - charge
fluorescent dyes used for
nuclear morphology
nucleotides have
covalent bonds
agarose, buffer, DNA and loading dye, assemble and load samples
A and T have
2 bonds
deciliate the cell by adding
OH groups, NH2 groups
ethidium bromide
fluorescent dye that lies betw the DNA bases
daltons are
base pairs or kilobases
A234/A260 show protein contamination when
carbon and hydrogens, has SH
3 kinds of microscopy
brightfield-wet mount
stereo-cult dish on stage
phase contrast-vary.deg. 1/4 wavelength from Zernike
DNA is twisted to the
left and has 2 backbones
A max
260 for nucleic acids
A vs. concentration
(direct linear proportion)
Beers-Lambert law
alkaline lysis proced
cells are pelleted and resuspended in a buffered medium, cells lysed w/SDS, RNase, NaOH, (SDS and NaOH solubilize the prots and elevated pH of NaOH degrades the RNA) alkaline condits denature chromosomal DNA and plasmid DNA separates the strands, salt concentration increases by adding potassium acetate-->chrom DNA, prots, SDS ppt while shorter plasmid renatures, ppted SDS, prots, chrom DNA centrifuged and plasmid in SN is ppted w/EtOH and dissolved in Tris/EDTA
number of codons
A320 should be
<5% A260
highly basic proteins
denaturing electrophoresis
prots denatured in urea, SDS, or a reagent that unfold tertiary struct
food is engulfed by
endocytosis-membrane surr. food and sequesters it in a vacuole
nucleus has a
fibrous matrix and various DNA binding prots that serve to regulate both transcription and DNA replicat
cooperative action of highly basic proteins known as histones organize nuclear DNA into
several chromosomes
2º structure
alpha helix and beta plated sheets are held by H bonds and can be a single polypeptide
A320 shows
light scattering properties
native electrophoresis
separates prots in buffered environmt maintains native charge and configuration
fluorescent dyes
coiled coils locked together and polymerize
can prevent annealing of the restriction cut sites and backbone repair by its treatmt
the logarithm to the base 10 of the reciprocal of the transmittance
DNA w/concentration of 50 ug/ml has
A260 of 1
coiled coil
amphipathic wraps around another amphipathic
A min
an instrument that provides a quantitative measuremt of the absorbed wavelengths
higher ratio for A260/A280 means
all prots have
1º and 2º structures
tetrahymena are
fresh water protozoan living in axenic culture
3º structures
3 D conformat due to folding of polypeptide and interactions of hydrophobic and hydrophilic
Euk cells have
complex cytoplasmic membrane systems like ER and membrane compartmts like mitochrondria
principle that charged molecules migrate in an electric field
cytoskeleton-netwk of interlock. structs consist of
microtubules, microfilaments, intermediate
hydrophobic aa's in helix spaced 3 or 4 posits apart in a sequence
indirect immunofluorescence
binding of secondary to primary enables us to see fluorescent image
1º struct
a.a in polypeptide chain w/peptide bonds
cytoskeleton contains many
basal bodies arranged in longitudinal row, each w/attached cilium, located near a complex called oral apparatus
mRNA-tRNA-prots (ribosomes)
globular prots are
tertiary structures
direct immunofluorescence using
DAPI(4'6 diamido 2 phenylindole)
-used on nonliving cells since it cannot penetrate the plasma membrane of living cells, therefore etOH is needed to fix the cells and then label them w/DAPI
transmitted light generates a
electric current when it strikes a phototube
1 or more nucleoli define
intense regions of RNA synthesis
photoelectric tube
is connected to a galvanometer and equates current w/% transmittance