DAT biology- microscopy and lab techniques

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restriction fragment length polymorphisms (RFLPs) and short tandem repeats (STRs) are used in ___ which is a technique that may be used in paternity and forensic cases

DNA fingerprinting

PCR can be carried out in a single container- list the components that container needs to contain in order for PCR to take place

DNA to be cloned, nucleotides, DNA, primers, heat-resistance DNA polymerase (Taq polymerase)

___ is a condition that results in a third copy of chromosome 21 and ___ allows for substantiation of its diagnosis

Down syndrome (trisomy 21), karyotyping

is SEM or TEM used to look at surfaces

SEM

describe the general idea of a pulse chase experiment for proteins

add radioactive aa during the pulse -> radioactive protein synthesis -> wash away radioactive aa -> add regular aa during the chase

southern and northern blotting tend to use ___ gel whereas western blotting tends to use ___

agarose, SDS-PAGE

what are 2 pep methods for selecting bacterial cells that have. undergone transformation in bacterial cloning

antibiotic resistance and color change

ELISA is based on the idea that a person will have ____ for a given diseases ___ of they have the disease or have been exposed to it

antibodies, antigens

___ are less common than sticky ends and they do not have unpaired nucleotides

blunt ends (harder to hybridize bc of the paired nucleotides)

what are some advantages of electron tomography?

can look at objects and their relative positions in 3D

what are hemocytometers

cell counting chambers

___ is the process where cell contents are separated into their fractions ( one part of a whole) by centrifugation

cell fractionation

gel electrophoresis separates macromolecular fragments on their ___ and ___

charge and size

colony forming units are based on the assumption that each viable cell initially plated gave rise to a ___

colony

___ are used to estimate the number of cells plated on a growth medium

colony forming units

what are some adv of confocal laser scanning optical microscopy

colorful, 2D image of living cells, view chromosomes during mitosis, overcomes fluorescence distortions (higher resolution)

what are some adv of fluorescence optical microscopy

colorful, 2D images of thin samples of living cells, inc brightness

___ is DNA made from RNA and to is used in microarrays and bacterial cloning

complementary DNA (cDNA)

___ are light microscopes that focus visible light to produce a 2D image of thin samples (single cell layers)

compound microscopes

___ is like SEM but the sample is frozen instead of dehydrated

cryo-scanning electron microscopy

in ___ only scattered light from the sample is transmitted to produce 2D images of unstained living cells

dark field optical microscopy

what are the 3 cyclical steps of PCR

denaturation, primer annealing, elongation

___ centrifugation separates cell contents in just 1 spin step creating multiple layers separated by density

density

____ is an older and more established method of DNA sequencing, while _____ is used more often now because it is quicker and cheaper

dideoxy chain termination (Sanger sequencing); next generation sequencing

what are the 2 most common method for DNA sequencing

dideoxycytidine chain termination (sanger sequencing) and next generation sequencing

in ___ cells are split open w a blender and the resulting homogenate is separated based on mass, density and or shape

differential centrifugation

what are the 3 germ layers from superficial to most deep

ectoderm, mesoderm, endoderm

what are 2 methods for automated cell counting

electrical resistance and flow cytometry

most viruses are so small that they mist be viewed using ___ microscopy

electron

___ integrates multiple TEM 2D images into a 3D model

electron tomography

___ microscopy allows for higher magnification than ___ microscopy

electron, optical

___ is a process where electricity is applied to cells creating temporary holes in the plasma membrane

electroporation

___ is a technology to determine if a specific antigen exists in a person, aiding in the diagnoses/exposure to certain diseases

enzyme-linked immunosorbent assay (ELISA)

bacterial cloning is an important technique to produce medicines bc ___ are cloned in ___

eukaryotic gene products, prokaryotic cells (insulin gene is obtained as cDNA from processed human mRNA

what is an advantage of dark field optical microscopy?

excellent contrast on living samples of unstained cells (black background)

microscope_____ adheres cells two microscope slides in their most lifelike state, and makes it easier for those cells to be stained

fixation, stained

in ___ cells pass through a very narrow tube and can be counted via detection by a laser beam

flow cytometry

___ is the emission of photons (light) from a particle that has absorbed light

fluorescence

___ provides a quantitive measure of the concentration of various ions, molecules. and gases in a cell

fluorescence lifetime imaging microscopy (FILM)

___ gives quantitive insights into cell dynamics bc bimolecular movements in live cells can be traced

fluorescence recovery after photobleaching (FRAP)

what are some disadvantages of fluorescence optical microscopy

fluorescence sometimes create distortions that reduce resolution

___ and ___ are optical microscope techniques that use laser light to produce 2D images of samples that have been tagged with fluorophores

fluorescences, confocal laser scanning

___ refers to the amount of time it takes for an excited molecule to release all its fluorescence

fluorescent lifetime

___ are fluorescent chemicals that will reemit light upon being excited by another light source

fluorophores

a ___ is a way to store all the DNA of an organisms genome

genomic library

___ studies all the genes of a genome by looking at their structure and function as well as their interactions with each other

genomics

what are some advantages of phase-contrast optical microscopes?

good resolution and contrast, can be used to observe thin samples of living cells- including their internal structure

what are some advantages of TEM?

high resolution 2D images of internal sample structures

what is an advantage of SEM

high resolution, 3D images of sample surfaces

what are some advantages of cryo-SEM?

high resolution, 3D images of sample surfaces, which are presented in a more natural form than SEM (due to freezing)

what are some disadvantages of phase contrast optical microscopes

ineffective on thick samples, halo effect around sample edges

what are some disadvantages of electron tomography?

it is costly and the extensive preparation kills all living cells

what are some disadvantages of TEM?

it is costly and the extensive sample preparation kills all living cells

what are some disadvantages of cryo-SEM?

it is costly and the fixation, staining, freezing kills the sample

what are some disadvantages of SEM

it is costly, and the fixation/staining/dehydration kills the sample

___ is the observation of chromosomes under a light microscope using staining

karyotyping

fixation and staining are associated w cells that have been _____

killed

compound light microscopes usually have different ___ which gives them the ability to make more resolute images than a stereo/dissection microscope

lens magnification

most optical microscope techniques can be used to view ___ samples of cells

living

describe the process of heat fixation:

living cells are placed on slide-> the slide is passed over a flame to kill/glue the cells to the slide -> stain is applied

what is an disadvantage of dark field optical microscopy?

low light intensity

centrifugation separates cell components through ___

mass, density and or shape

karyotyping is performed during ___

metaphase

a ___ is a chip containing thousands of probes that are complementary to a certain sequence

microarray

____ cells can give rise to some of the 3 germ layers but not all so their are the most differentiated and cannot develop an entire organism

multipotent

can electrons microscopy be used to look at living specimens?

no due to fixation and staining

___ is an electrophoresis technique for separating RNA fragments and is used ___ probes

northern blotting, RNA

arrange the following organelles from most to least dense: ER, ribosomes , mito, nuclei, chloroplasts

nuclei > mito/chloroplast > ER > fragments > ribosomes

as cells show electrical resistance and impede conductance the ___ in a solution can be estimated by observing the flow of electricity

number of cells

a karyotype shows both the ___ of chromosomes and their ___

number, physical appearances

a ___ occurs when there is a black of nucleotides that are inverted mirrors of each other

palindromic sequence

___ are optical microscopes that use light phase changes and contrast to produce 2D image of thin sample

phase-contrast microscopes

___ catalyzes phosphodiester bonds bw the ends of DNA restriction fragments (used heavily in genomic libraries and bacterial cloning)

plasmids

a ___ cell can give rise to an of the 3 germ layer but they cannot develop and entire organism bc they can't develop extra embryonic tissue

pluripotent

the ___ is an automated biotechnology process that can quickly create millions of copies of DNA and it requires no cell

polymerase chain reaction (PCR)

a ___ is a fluorescent or radioactively labeled tool that allows scientists to identify a specific sequence within a large sample

probe

___ corresponds to a eukaryotic gene with all intros removed (used in microarrays and bacterial cloning)

processed mRNA

centrifugation can be used to separate ___ based on solubility

proteins (insoluble proteins pellet out while the soluble proteins remain in the supernatant)

___ experiments allow for the visualization/tracking of molecules of interest throughout a cell

pulse chase

___ is produced when DNA fragments from different sources are joined together

recombinant DNA

knockout mice have a selected gene of interest knocked out by ____

recombinant DNA technology

what are some disadvantages of confocal laser scanning optical microscopy

reduced light intensity and longer illumination times than fluorescences optical microscopy

___ is the process of reverting a multipoint somatic cell to totipotency such that it can develop into another identical animal

reproductive cloning

the DNA fragments that get incorporated into recombinant DNA are produce by ___ which tend to cut DNA at palindromic sequences to produce ___ ends

restriction enzymes, sticky or blunt

___ are unique lengths of DNA that result from restriction enzymes allowing the comparison bw individuals

restriction fragment length polymorphisms (RFLPs)

___ produces complementary DNA (cDNA) from mRNA, and it is relied upon for microarrays and bacterial cloning

reverse transcriptase

what are the 3 components of a chromatography apparatus

sample, mobile phase, stationary phase

___ captures electrons that are scattered by atoms found on the surface of dehydrated samples

scanning electron microscopy (SEM)

in centrifugation the densest and most compact particles will ___ to the bottom of the tube first becoming pressed together as a ____ (precipitate)

sediment, pellet (top liquid is the supernatant)

a ___ is a group of nucleotides that repeats again and again in a stretch of DNA

short tandem repeats (STRs)

for the most part the human genome is the same with slight differences in the sequence every 1000 nucleotides (called ___) which serve as markers for genes that caused disease

single nucleotide polymorphisms (SNPs)

in gel electrophoresis the largest or smallest fragments travel the furthest

smallest

___ is an electrophoresis technique for separating RNA fragments and it uses probes

southern blotting, DNA

___ is the process of adding color to cells which allows them to be viewed more easily under microscope

staining

___ are light microscopes that focus visible light to produce a 2D image of a staples surface

stereo-microscopes (dissection microscope)

what is an advantage of stereo-microscopes/dissection microscopes

they are light microscopes that can be used to view living samples

what is a disadvantage of stereo/dissection microscopes

they are light microscopes that have low resolution

what is an advantage of compound light microscopes

they can be used to view 2D images of living samples (1 cell thick)

what are some disadvantages of compound light microscopes?

they only view samples that are 1 cell thick and they have poor contrast, which means some samples my need to be fixed/stained/killed

___ cells have the ability to produce extra embryonic membranes and ultimately an entire organism

totipotent

___ is a process that occurs when a cells genome is changed by the addition of DNA that was once floating freely in the environment

transformation

competent bacterial cells can undergo ___ and they can be made competent through ___

transformation, electroporation

___ have genes from different species in order to study gene effects and or mass produce medications

transgenic animals

___ capture electrons that are transmitted through a thin slice of a sample

transmission electron microscopy (TEM)

sticky ends have ___ which makes it easy for complementary sticky ends to hybridize

unpaired nucleotides (complimentary sticky ends are made by the same restriction enzymes)

what are some strategies to reduce the halo effect of phase contrast optical microscopy

using phase plates to reduce the phase shift, use thinner samples

in electron microscopy electrons are shot through a ___ at a sample which has been mixed and metal coated (cells are dead)

vacuum (prevents electrons from deviating in path)

a ___ is a piece of DNA (such as a plasmid) that can be taken up by complement cells

vector

___ is an electrophoresis technique for separating proteins and it uses ___ as the probes

western blotting, primary and secondary antibodies


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