Biotechnology and Microscopy Flashcards
Uses visible light and optical lenses to magnify and view a sample
optical microscopy
Uses a focused beam of electrons to magnify and view a sample
electron microscopy
Optical microscopy allows us to see most ___ and __ cells, most ___, and ___. However, to see ___, ___, and -___ electron microscopy is needed. Some long ___ are visible with the naked eye
animal plant, bacteria, organelles, viruses, ribosomes, proteins, neurons
Compound microscope is a type of ___ microscope in which visible light is focused on a thin -_ of the sample to view it in ___. It is used for the observation of __, __ and ___. ___ can be used for enhanced viewing, While very ___ single cell layers do not need this, ___ samples require it, and in the process will kill the sample
light, slice, 2D, cells, tissues, organisms, staining, thin, thicker
fluorescence microscopy uses a __ __ to tag certain structures. It can be useful to locate an ___ or where __ ___ is within a cell. This can be used on ____ cells in real time, for example to look at ___ during mitosis
fluorescent marker, organelle, protein expression, living, chromosomes
In scanning electron microscopy, a ___ image of the sample’s ____ is produced in very high resolution, allowing us to see the _____ and ____ of small structures. It is ideal for viewing the __ ___ of cells, tissues and molecules, but the sample must first be ____ and ___ before viewing, which will kill it
3D, surface, texture, shape, external surface, dehydrated coated
Transmission electron microscopy, uses an electron beam passed through a very ___ section of sample to produce a high magnification ___ image. It allows for high resolution viewing of ____ ___, and even inside ____. It has the highest ___ of all microscopes. However, the preparation of the sample is ___, ____ and kills it
thin, 2D, internal structures, organelles, magnification, time-consuming, expensive
Using a differential centrifuguation to separate a cell’s contents based on density and size
cell fractionation
In cell fractionation, first _____ is needed, where cells are broken apart with the cell contents without a _____. Then the homgenate is spun at a ___ speed which creates a dense ___ ____ of ___. Then the layer is removed and the sample is spun on ___ speed producing a new pellet layer of ____ and ____. This repeats, and until the smallest cell components like the ___ and ___ remain as the pellet layer
homogenization, membrane, low, pellet layer, nuclei, medium, mitochondria, chloroplasts, ribosomes, viruses
Transfer of genes from generation to the next (e.g. sexual / asexual reproduction, mitosis)
vertical gene transfer
transfer of genes between different organisms
horizontal gene transfer
___ is a type of horizontal gene transfer in which DNA is directly transferred via a biological bridge between two organisms. This occurs between ____ that have a ____, which connects the cytoplasm of one bacteria to another
conjugation, bacteria, pilus
A type of horizontal gene transfer in which DNA is introduced into a genome by a virus
transduction
___ is when a cell absorbs DNA from the surroundings and incorporates it into their DNA. This can be conditioned via ___ ___ or _____
transformation, heat shocking, electroporating
DNA containing different segments from multiple sources
recombinant DNA
Recombinant DNA technology uses __ __ to cut up sequence specific sites called ___ ___, which are nucleic acid sequences that read in both the __ and __ direction. Restriction enzymes can produce ___ ends where there is overhang of nucleotides, which is more ___, and ___ ends, where there is no overhang
restriction enzymes, palindromic sequences, 5’->3’, 3’->5’, sticky, common, blunt
in recombinant DNA technology, if the ___ restriction enzyme is used to cut pieces of different sources of DNA, the ___ __ ___ of two DNA pieces can bind, creating a DNA molecule from multiple sources. Incubation with __ ___ will seal the phosphodiester backbone
same, unpaired sticky ends, DNA ligase
Creating a map of known restriction enzyme cut sites within a sequence of DNA to know where to cut and what genes are nearby
restriction mapping
the location of __ ___ on human DNA varies between individuals, meaning enzymes will create unique fragments of different ___ based on the individual. This concept is referred to as ___ __ __ and is useful in __ ___, when a DNA in a crime scene can be used to match up to an individual
restriction sites, sizes, restriction fragment polymorphisms, DNA fingerprinting
__ ___ __ are single nucleotide differences in the human genome, one in roughly every ______ nucleotides. This may be found near _____ ___, and thus can be used as genetic markers for _____ to certain diseases
single nucleotide polymorphisms, 1000-2000, disease-associated alleles, susceptibility
__ ___ can be used to separate DNA/RNA/proteins based on __ and __. It is made of a gel medium soaked in a _ ____ ___ solution hooked up to a machine that provides a negative charge on one end and ___ charge on the other
gel electrophoresis, charge, size, electrically conductive buffer, positively
The sample used in gel electrophoresis is loaded in the ___ at the ___ end. Then a ___ is provided by the machine, and DNA molecules which are _____ charged, will move towards the ____ side. The ___ DNA molecules move further than the ___ DNA molecules
wells, negative, current, negatively, positive, shorter, larger
After electrophoresis, DNA can be ____ or probed using a _____ labelled __ strand nucleic acid to identify the location of the specific sequence
sequenced, radioactively, single
Gel electrophoresis is similar but because they have a strong ___ structure, ___ must be added prior to loading which ___ the protein into a linear polypeptide chain and adds a uniformly _____ charge proportional to the size of the molecule
folded, SDS, denatures, negative
When nucleic acids of one strand form base pairs with complementary nucleic acids on a different strand
nucleic acid hybridization
A DNA probe is often labelled ___ or ___, then ___ so that it is single stranded. Then the target strand is also denatured, and if it is present, the probe can ____ to it. The detectable tag can be used to ____ the target strand. This technique is used in _____ ____
radioactively, fluorescently, denatured, hybridize, locate, in-situ hybridization
In situ hybridization, the probe is labelled with a ___ dye, and hybridized with the ___ of interest, testing the location and presence of ___ ___ in an organism.
fluorescent, mRNA, gene expression
DNA or ___ ___ ___ sequencing is used to determine the number of __ __ in a DNA or RNA molecule and their ____
dideoxy chain termination, base pairs, sequence,
In DNA sequencing, first the desired DNA strand is ____ into a single strand form. Then it is mixed with a ____ which provides the _____ necessary to begin DNA synthesis. Finally the sample is incubated with ___, ___, and fluorescently tagged _____
denatured, primer, 3’OH, DNA polymerase, deoxyribonucleotides, dideoxy-ribonucleotides
The _____ unlike deoxyribonucleotides, lack a ____ and thus cannot form a ____ bond with another nucleotide. This is the ___ ___ nucleotide
dideoxyribonucleotides, 3’OH, phosphodiester, replication terminating
In DNA sequencing, hundreds of DNA synthases continue, each ___ stopping at a specific __ ___. Ultimately, a set of labelled strands of every possible ___ is created. Therefore, we have a fluorescent indicator on every single ____
randomly, fluorescent ddNTP, length, nucleotide
In DNA sequencing, after the strands are made, the labelled strands are separated via ___ ___ ___ from the shortest to longest strand. This while mean the ____ strand with only ___ nucleotide will pass through first. Eventually it is possible to get the ___ of the nucleotides from technology that detects the order of the __ __
capillary gel electrophoresis, first, 1, order, fluorescent colours
In reverse transcription an enzyme called ____ ____ is used to synthesize DNA molecules off an ___ template. This is makes a complementary _____ with no ___. Some viruses like __ and ___ use this to replicate their genome and proliferate in the host
reverse transcriptase, cRNA, cDNA, introns, HIV, hepatitis B
Reverse transcriptional is used to make __ __ in bacteria. the cDNA is needed because prokaryotic RNA does not contain ___, and thus have no mechanism to remove them. The cDNA allows for the desired gene to be efficiently transcribed and ___ after insertion. mRNA itself cannot be used because it is unstable and ____
recombinant DNA, introns, translated, short-lived
Technique used for the amplification of DNA
polymerase chain reaction
In PCR, a double stranded DNA is first ___ to separate into strands, thus requiring a __ ___ ___. In ____ as the temperature cools down, the primers can attach to separate strands. In ____, the temperature is raised again, and heat resistant ___ synthesizes complementary strands. This is usually done by ___ ___ which is more stable under heat. This replication process occurs for ___ strands of DNA, multiple times, ___ increasing the number of DNA molecules
heated, heat resistant polymerase, annealing, primers, elongation, polymerase, prokaryotic polymerase, both, exponentially
DNA microarray assay is used to monitor the expression of ___ groups of ___ across the entire genome. This is useful for seeing which genes are ___ in different tissues or at different stages of ____
large, genes, transcribed, development
DNA microassay contains many wells containing __ ___ pieces of DNA, all unique. Then ____ ___ made from the ___ of target cells are loaded into the wells to hybridize. This allows us to see what genes are expressed in ___ vs ___ cells, as the cells containing the genes of interest will be ___ labelled. It can also be used to look at different __ of cells
short single, fluorescent cDNA, mRNA, healthy, cancerous, fluorescently, types
blotting technique used to look for specific genes in DNA fragments
southern blot
blotting technique used to look for specific genes in RNA molecules
northern blotting
Blotting technique used to identify proteins
western blotting
In southern blotting, the DNA is first extracted from a biological sample and ___ via __ ___. Then the DNA fragments are separated according to their ___ via ___ ___. The fragments are then transferred to ____ ___ ___ by blotting paper. This paper is very thin, helping in visualization, and easily allows for a DNA probe with a ____ ___ or ____ tag to bind because it is ___. Any DNA fragments containing __ ___ can be detected
cut, restriction enzymes, size, gel electrophoresis, nitrocellulose filter paper, radioactive, fluorescent, chemical, porous, complementary sequences
allows for the visual identification of proteins
immunofluorescent staining
In immunofluorescent staining a __ ___ binds to the protein of interest. Then a ___ __ containing a ___ __ will bind to the secondary antibody. Using microscopy, the tag can be visually ___ to detect the protein of interest in a ___ sample
primary antibody, secondary antibody, fluorescent tag, located, live
in vivo mutagenesis helps determine the ____ of a gene. This introduces specific ___ into a gene, and observe ____ differences. These differences may be a function of a missing __ ___. A frequently used example is ___ ___
function, mutation, phenotypic. normal protein, knockout mice
similar process to in-vivo mutagenesis but only studies the effects of the mutation outside of a living organism
in-vitro mutagenesis
__ _____ analyzes genomic sequences to identify ___ ___ and their functions. it utilizes ___ __ to compare known sequences
genome annotation, protein-coding regions, computer databases
The introduction of genes into an afflicted individual for therapeutic purposes
gene therapy
in gene therapy, first the cell with the genetic disease is ____, then loaded into a __ ___, which will infect the cell with the good genes. These cells then are ___ into the person
isolated, retroviral vector, reinjected
Animals which have had a gene introduced from the genome of another individual - often another species
transgenic animals
Transgenic animals can be used to study the __ of genes, or in drug testing to measure __ ___ and replicate __.
function, drug efficacy, diseases
In transgenic animals, the cells of both animals of interest are first ___. Then the gene of interest to be insert is taken and placed ____ to the gene that codes for what will express the gene of interest. Then the nucleus is injected into an ____, and then implanted
isolated, adjacent, embryo
collection of cloned DNA pieces from a genome; used to locate a gene of interest
genomic library
To form the genomic library, first the genome of interest is ___, and then it is cut leaving __ ___ with a __ ___. then a _____ or ____ DNA is cut with the same restriction enzymes. These plasmids are designed to include an __ ____ gene. Then the DNA and plasmids are sealed together using __ __.
isolated, sticky ends, restriction enzymes, plasmids, circular, antibiotic resistance, DNA ligase
In forming a genomic library, the ___ DNA is inserted into a bacteria by ____. This can be done via ___ __ and ___, or by ____. Then the bacteria are allowed to undergo repeated ___ __ to produce a population of __ ___ cells with the recombinant DNA. The genome library can now be ___ again for use or experimentation
recombinant, transformation, heat shock, CaCl2, electroporation, cell divisions, recombinant plasmid, isolated
When a cell is exposed to a brief electrical impulse, creating temporary pores in the plasma membrane
electroporation
When the temperature is increased then rapidly cooled, increasing the membrane permeability
heat shock
The plasmids used in genomic libraries contain __ __ genes, to do a test. All the bacteria are treated with an ____, and the bacteria that don’t survive did not successfully accept the ___, whereas the ones that did have are called the __ ___
antibiotic resistance, antibiotic, plasmid, recombinant
producing an organism that is genetically identical to the parent
cloning
In cloning first, the ____ from the ___ cell of an organism is isolated. Then, an ___ ___ is removed from the female and it’s nucleus is removed, producing an ___ egg. Then, ______ ____allows for the nucleus and egg to merge. This will form an ___ that will undergo cell division. This is ___ in the uterus of a surrogate mother, and the baby clone is birthed
nucleus, somatic, unfertilized egg, enucleated, electric shock, embryo, implanted
The ___ and the ___ ___ contributes no genetic information to the clone. The first clone made was of a ___
egg, surrogate mother, sheep
Pasteur’s __ ___ flask experiment proved that ___ ___ ___ was invalid, and that life could only be created from ___ organisms
swan neck, spontaneous life generation, existing
In pasteur’s experiment, the a broth was kept in a __ ___ flask, which prevented microorganisms in the __ from being able the enter the solution. Then the solution was __ to kill all the ___. When the curved neck was remained on, there were ___ microorganisms, while if the neck was kept off, the microorganisms grew due to entering from the air.
curved neck, air, boiled, microorganisms no
In Griffith’s experiment, two strains of ____ were injected into mice. The ___ strain lacked a protective capsule and there were destroyed by the mice’s immune system before it could kill it, therefore being ____. The ___ strain had a protective capsule that shielded it from the immune system, causing it to be ____.
bacteria, rough, non-virulent, smooth, virulent
In Griffith’s experiment, when S bacteria were ___ and injected, it would not harm the mouse. However, if it was heat killed and then added to a solution of living ___ bacteria and then injected, this would kill the mouse. This is because the R bacteria ____ the S ____, and this allowed it to produce the __ __ that shielded it from the immune system. This led to the discovery of the term _____
heat-killed, R, absorbed, DNA, protective capsule, transformation
In Avery-Macleod-MacCarty’s experiment, they expanded on Griffith’s experiment by heat killing the S cells, removing the ___ and ___, and separating the solution. In each separate solution they added digestive enzymes like ___, ___ and ____ to get rid of proteins, RNA, and DNA. When ____ was added to the S bacteria, the R cells were not transformed. This led to the conclusion that ___ was responsible for coding for the protective capsule
lipids, sugars, protease, RNase, DNase, DNase, DNA
A virus that infects bacteria
bacteriophage
Hershey and Chase radioactively labelled a bacteriophage called ____ ___with either a ____ to detect proteins, or with ____ to detect DNA. Then they let the bacteriophage infect a bacteria, and observed that only the radioactive ___ appeared inside the bacteria, confirming that DNA was the ___ ___ of the virus. This is because when a virus injects its information into a cell, it discards its ___ ___ outside of the cell which ___
phage T2, sulfur, phosphorous, DNA, genetic material, protein coat, dissolves
in meselson and stahl’s experiment, they grew ___ in a medium with nucleotides containing ____ a heavy isotope of nitrogen. The bacteria would then form ___ with this isotope. Then the bacteria were transferred to a medium with ___, where they would incorporate the nucleotides with the regular isotope. As they replicated,____ synthesized strands would have the lighter isotope while the ___ strands would have the heavier isotope
e. coli, n15, DNA, n14, newly, original
In meselson and stahl’s experiment, when the DNA was ____, the resulting DNA was not as __ as the original DNA but not as ___ as the 14N DNA, implying ___ ___. When a second round of replication occurred, there was a DNA strand with only ____, and a ____ combination DNA, further proving semi conservative replication
dense, light, semiconservative, N14, N14-N15
In Gurdon’s _ ___ experiment, an ____ cell was taken from a frog and it’s ___ was removed. Then this cell was fused with a ___ ____ cell. The egg was then stimulated to undergo ____ _____, which gave rise to a new frog. This proved that fully ___ cells do not lose their genetic information, they retain the full ___
nuclear transfer, intestinal, nucleus, enucleated egg, embryonic development, differentiated, genome
