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bio research Flashcards

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1
Q

ELISA TECH is used to detect?

A

antigens (proteins and cytokines) and specific antibodies

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2
Q

if they are testing for an antigen, what do they use?

A

they coat the wells with antibody specific to the antigen . then they wash it off after binding occurs. then a second antibody is introduced and linked to an detection enzyme.

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3
Q

if they are testing for an antibody, what do they use?

A

antigen goes directly to the wells to detect the antibody and then it is ,mixed with enzyme linked antibodies

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4
Q

elisa immunoassays vs radioimmune assays

A

elisa will have a color change whena presence of the diesired is present and deals with enzyme linked antiobodies

RIA is commonly used more for detecting certain hormones in patients serum… deals with mixture of radiolabeled antigen and antibody to cause a radioactivity that is compared to the standard curve

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5
Q

electrophoresis

A

separates dna rna or proteins by size

gel can be agarose or acymilide

direction includes the neg going to the pos side

thicker the gel, the smaller the pores so the small things are able to travel faster than the big ones

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6
Q

what is blotting

A

once electrophoresis is complete, the desired dna rna or protein will be isolated and placed on a nitrocellulose membrane and begin probing the fragment or protein

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7
Q

what does snow drop stand for

A

diff blotting

SOUTH = DNA
NORTH = RNA
WEST = PROTEIN
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8
Q

recombinant protein

A

basically when u clones genes of dna from diff organisms and trsnacribe/ translate to make a combination of it which can t hen help to cure diseases or work as treatment

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9
Q

what is the significance of the plasmid?

A

u place the desired gene into the plasmid so that it can grow with the bacteria

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10
Q

although plasmids are good, prokaryotes lack introns so how can plasmids process the exact gene that u desire

A

use complentart dna with the help of reverse transcriptase to change the rna into complentary dna

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11
Q

how are eukaryotic plasmids introduced to mammalian host cells

A

via transfection using calcium phosphate or plasmid packaging in liposomes …

Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells

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12
Q

what are the agents involved for eukaryotic plasmids?

A

puromycin or neomycin

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13
Q

why is polymerase chain reaction important

A

allows amplification and analysis of very small smaples of dna

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14
Q

what are the different things that are used for pcr

A

fingerprints , cloning genes, infectious diseases, hereditary info ect

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15
Q

what does and doesn’t the reverse transcriptase polymerase chain reaction do (RT-PCR) do ?

A

it checks the relative expression of specific gene products like mrna transcription but not the actual

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16
Q

how does reverse transcription pcr work

A

take the gene and isolate it. then use reverse to encode into the dna … then with help of primers to identify the complentary dna strand of the mrna

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17
Q

what is quantitative polymerase chain (qPCR), what is it performed on, and how is it detected?

A

finds the absolute number of the copies or the relative number normalized to the control.

can be performed on either dna or cdna

detected through fluorescent that will allow dna to bind or through flurescent probe

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18
Q

dna sequencing

A

is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine.

helps with studying health and disease

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19
Q

ribose vs deoxyribose 2nd carbon

A

hydroxyl on second carbon of ribose

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20
Q

in dna sequencing technique, why is the third hydroxyl group missing

A

to prevent elongation of dna strand to occur

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21
Q

describe the dna sequencing

A
  1. get sample dna and denature it
  2. mix in 4 separate test tubes aand add a different dideoxybase to each tube
  3. let replication occur and see different fragments through electrophoresis.

the bottom starands are the ones farthest from the wells
3.

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22
Q

genomic sequencing and what two approaches it can use?

A

help to tests cancer by generating

a genetic map linkage with several hundred markers per chromosome where it has a large library of chromosomes but gets smaller and smaller as more cloning takes place

shotgun that skips the gene mapping and cuts the chromosomes into fragments

23
Q

to convert lactic acid back to pyruvate, u need what molecule

A

o2

24
Q

polymorphisms

A

stretches of repeteitive highly variable dna

2-100 bp

25
Q

two types of analysis used for dna fingerprinting

A

restriction fragment length polymorphism
—- includes using restriction endonucleases to cut the polymorphisms into dna fragments that vary in size which is unique to humans

separated using gel electrophoresis and dna south blott

short tandem repeat
uses pcr to amplify polymorhisns that are found in non coding introns

26
Q

exome and targeted sequencing benefit

A

only target genes of interest through sequencing only the exons

27
Q

flurorescence in situ hybridization (FISH)

A

uses fluorescent probes to locate the positions of specific dna sequences which helps to see presence or absence of specific dna sequences

28
Q

microarrays

A

used to study relative rna amounts btween two samples or to compare one to the standard reference.

29
Q

steps to microarrays

A

place fluoresxent dye

place on chip that has the binding site for every gene

30
Q

what does the chip do in micro array?

A

has the binding site for every gene that acts as a probe to determine transcript levels in tissue samples

31
Q

when it comes to the respire sys, what relaxies it

A

sypathet

32
Q

in situ hybridization (ISH)

A

similar to fish but examines examines gene of interest in a tissue that is fixed to keep transcripts in place

33
Q

Immunohistochemistry (IHC)

A

needs an antibody that is recognized by a second antibody which is either linked to an enzyme or a fluroscent molecule (staining)

used to identify breast tumors

34
Q

flow cytometry

A

single cells stained for certain protein markers using specific antibodies linked to fluorescent tags that can emit light

35
Q

what does the scattered and emitted light show in flow cytometry?

A

gives info on cell size and how many cells in the sample express each of the markers that were labeled

36
Q

describe immunoprecipitations

A

commonly used to find protein binding partners

includes cell lysates, antibody specific for protein of interest

37
Q

what happens in immunoprecipitations

A

antibody has bead that its covalently attached to which can be pulled out of soln or precipitated

use a lysate to wash the undesired proteins away

38
Q

mass spec vs western blot when it comes to protein detection

A

use western when u have an idea of the protein u are looking for. if u don’t, use mass spec

39
Q

subcellular location

A

find the location of the protein by using an expression vector tgat traces the protein with green flurescence

40
Q

why is altering gene expression useful

A

it can help determine function of protein by knocking out/in genes , exciting or inhibiting , or looking at the phenotype

41
Q

in vitro mutagenesis

A

clone a gene, mutate it, place back into a cell to see alter the gene fuction that u are studying

42
Q

what are stem cells and do they use mitosis or meiosis

A

undifferentiated cells that differentiate to become other cell types ..

replicate by mitosis

43
Q

what are pluripotent cells and give an ex

A

able to differentiate into any of the three germ layers

ex: embryonic cells

44
Q

what are adult stem cells used for and give 3 ex of sources

A

tissue repair and regeneration

ex: bone marrow, adipose tissue, blood

45
Q

sugars have l or d

A

carbohydrates (sugars are deliscious)

46
Q

difference in probes btween western vs northern and southern

A

antibodies are used in northern and southern for probes… in western they nucleic acids

47
Q

gene therapy

A

genetoic disorder is treated bhy introducing to a cell

uses genetically modified virus… retro and adeno viruses

48
Q

affinity chromatography

A

biochem mixture that ius based on interactions:

purify proteins

antigen and antibody
enzyme and substare
receptor and ligand
protein and nucleic acid

49
Q

turning region

A

loose unstructured region that is not in the alpha or beta structures

50
Q

what is wrong about glycine?

A

its so smalls so it disrupts secondary structure

51
Q

why is a third ionization energy higher than first and second?

A

3rd electron is being removed from a complete full subshell

52
Q

which aa have chiral carbons in side chains?

A

threonine and isoleucine

53
Q

bases can cut enzymic activity ?

A

bases can cleave peptide residues

MCAT PREP (14 decks)

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