Molecular Diagnostics exam Flashcards by amanda gonzalez

In end-point PCR gel, what are the fragments that migrate past the smallest standard?

Primers and primer-dimers

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During the transfer of DNA for Southern blotting, larger fragments of DNA do what?

Transfer more slowly than smaller fragments

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In the denaturation step of the polymerase Chain Reaction what happens to the dsDNA?

Gets separated into ssDNA

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What are the steps to southern blot procedure?

Restricted enzyme
electrophoresis
denaturation
probe hybridization

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What is the signficance of the observation of circulating mRNA versus the observation of circulating DNA?

The presence of circulating mRNA indicates that a message has been transcribed and will likely result in the synthesis of new protein

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Which reporter unit in real-time PCR is designed in a hair-pin structure?

Molecular Beacon

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Fluorescent dye specific to dsDNA?

SYBR Green

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This method exploits the natural 5’ to 3’ exonuclease activity of Taq polymerase to generate signal?

TaqMan Probe

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This method utilizes specific probes, one with a 3’ fluorophore (acceptor) and the other with a 5’ catalyst for the flurorescence (donor), that bind to adjacent targets?

FRET (fluorescent resonance energy transfer

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The conversion of mRNA nucleotide sequences and the transfer RNA tRNA attached amino acids into a polypeptie is referred to as what?

translation

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The copying of one strand of DNA into RNA by a process similar to that of DNA replication?

Transcription

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What are UV spectrophotometry, microfluidics technology, and gel electrophoresis used for?

quantitating DNA concentration

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What method uses antibodies bound to the surface of a microtiter well to detect the presence of human papilloma virus?

Hybrid capture assay

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Singnal amplification system based on th eproprietary cleavase enzyme?

Cleavage-based amplification

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Replication of a DNA or RNA target through an intermediate RNA product?

Nucleic Acid sequence-bases amplification (NASBA)

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What determines the specificity in terms of the gene that is detected in the Southern Blot method?

Labeled probe

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The steps to DNA isolation?

Cell lysis
precipitation
protein digestion

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nitrogens with a single ring structure?

Cystosine and Thymine (pyrimadines)

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nitrogens with a double ring structure?

Adanine and guanine (purines)

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This enzymes breaks the sugar-phosphate backbone of DNA?

Endonuclease

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These are endonulceases that recognize specific base sequences and break or restrict the DNA?

Restriction enzymes

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Denaturation temp?
annealing temp?
DNA extension temp?

Denaturation 94-96 degrees
annealing 50-70 degrees
DNA extension 72 degrees

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What is DNA—>Transcription—>RNA—>translation—->protein?

Central Dogma of molecular biology

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Nucleic acids migrate toward which pole in electrophoresis?

anode

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What dimultaneously detects two or more different targets from one polymerase chain reaction tube?

multiplex pcr

strand of the parent double helix that is read 5' to 3' and replicated discontinuously?

lagging strand

strand of parent double helix that is read 3' to 5' and replicated continuously?

leading strand

polyadenylic acid at the 3' terminus of mRNA

poly A tail

Intermediate products of DNA synthesis on the lagging strand

Okazaki fragments

DNA that is generated from RNA prior to amplification is called what?

Complementary DNA (cDNA)

These occur only in microorganisms, recognize sequences of 4-15 bp in length, and cleave phophodiester bonds

restriction endonucleases

The most commonly used polymerasein the PCR procedure?

Taq polymerase

Stringent hybridization conditions for southern blotting?

High temp, low salt concentration

What wavelength of light is protein absorbed in our nucleic acid suspension?

280 nm

What enzyme generates single stranded DNA from ribosomal or messenger RNA?

DNA polymerase

Which enzyme would you use to join together 2 seperate DNA fragments?

DNA Ligase

What are histone modification, DNA methylation and Gene splicing?

forms of epigenetic changes

A region of DNA present in a mature strand of mRNA and can be translated into protein?

Exon

Non coding sequences in DNA that interrupt coding sequences

Intron

Helicase, DNA template and DNA polymerasea are required for what?

DNA replication

Cytosine and Guanine have how many hydrogen bonds?

Adenine and thymine have how many hydrogen bonds?

The RNA processing sections of nucleotides are removed by:

splicing

A chromosome that has the centromere at the end of the chromosome is called:

telocentric

A chromosome that has the centromere in the middle of the chromosome

metacentric

A chromosome the entire length of which acts as a centromere

holocentric

In DNA the absorptivity constant is what?

50 ug/mL

In RNA the absorptivity constant is what?

40 ug/mL

phenomenon associated with tri-nucleotide repeat disorders of earlier onset and increasing severity of disease with successive generations?

anticipation

Method of DNA sequencing where dideocxy NTPs are incorporated into growing oligonucleotide chains, resultig in termination ddNTP's

Dideoxy (Sanger) sequencing

consisting of many clones, often reference to lymphoid cells

polyclonal

Individual with 2 abnormal alleles, each with a different polymorphism or mutation.

Compound heterozygote

mutation that is the change of a single base?

point mutation

process of forming a double stranded molecule from a single stranded probe and a single stranded nucleic acid target based on complementary?

hybridization

gene constitutively expressed inmost cells that can serve as an internal control for reverese transcriptase PCR?

housekeeping gene

In real-time PCR, the cycle where the exponential phase of an amplification curve begins?

crossing point

proofreads newly synthesized DNA and breaks the phosphodiester bond to replace it with the correct one?

exonulease

adaptors that link condons in mRNA with amino acids?

tRNA

an isothermal amplification reaction using nick translation and strand displacement...proceeds at 1 temperature

stand displacement amplification

Target DNA is released from the cells bound to ssRNA probes. Used in the detection of HPV.

Hybrid capture signal amplification

Single strand of a DNA double helix that is not used as a template for messenger RNA synthesis

sense strand

single strand of a DNA double helix that is used as a template for messenger RNA synthesis

anti-sense strand

breakage and fusion of separate chromosomes

translocation

alteration in the nucleotide sequence that does not change the amino acid sequence

silent

alteration in the nucleotide sequence that results in the substitution of an amino acid with one of similar properties

conservative

alteration in the nuleotide sequence that results in the substitution of an amino acid with on of dissimilar properties

non-conservative or missense

alteration in the nucleotide sequence that results in the substitution of a termination codon for an amino acid codon

non sense

deletion or insertion of other than a multiple of three base pairs in coding sequence

frameshift

method used to detect sequence alterations in DNA through differences in secondary structure due to differences in the nucleotide sequence?

Single strand conformation polymorphism

polymorphism detection technique using immobilized target and sequence specific probes

allele-specific oligomer hybridization

detection of point mutations or polymorphisms in DNA with primers designed so that the 3' most base of the primer hybridies to the test nucleotide position in the template?

sequence-specific primer

method using time of flight?

MALDI-TOF

genetic locus that differs between two individuals?

informative locus

< 1 % of the population have this

mutation

>1% of the population have this

polymorphism

a procedure that identifies specific nucleotide sequences localized within cells in a tissue section?

FISH | in situ hybridization

What molecular mechanism accounts from the emergence of methacillian resistance to staph aureus?

mutation of the Meca gene in the DNA that alters penicillin binding protein.

Micro satellites that repeat 2-10 bps are also known as what?

STRs

mini satellites that repeat 40-500 bp in length are called what?

VNTP's | variable number of tandem repeats

One base pair variation from a reference DNA sequence

SNP | single nucleotide polymorphism

biological properties of an organism?

phenotype

genetic DNA composition of an organism

genotype

reference size standards used in STR analysis representing all possible products (alleles) for a given locus?

allelic ladders

genetically linked set of alleles that are inherited together?

haplotype

the donor organ and the recipient are genetically different?

allograft

the level of detail with which an allele is determined?

resolution

What are the most important polymorphic loci in the MHC?

HLA-B | HLA-DRB

Chemical sequencing method based on controlled breakage of DNA?

Maxam-Gilbert method

Each nucleotide is added in turn only 1 in 4 will generate a light signal the remaining nucleotides are removed enzymatically the light signal is recorded on a pyrogram

pyrosequencing

replication of a DNA or RNA target through a intermediate RNA product

transcription-mediated amplification

inheritance pattern where the child of an affected and an unaffected carrier has a 50% probability of being affected and where the child of two affected individuals has a 100% probability of being affected Cystic fibrosis and hemochromatosis

autosomal-recessive

inheritance pattern where the child of an affected and an unaffected individual has a 50% probability of being affected Factor V Ligand and Huntingtons Disease

autosomal dominant

Having genetic components located on the X or Y chromosome | hemophilia

sex-linked

The frequency of expression of a disease phenotype in individuals with a gene lesion

penetrance

mutated gene that promotes proliferation and survival of cancer cells

oncogene

self directed cell death

apoptosis

the normal intrachromosomal rearrangements in B and T lymphocytes as well as the abnormal interchromsomal translocation that can occur in any cell type

V(D)J recombination