Genetic Variations & Molecular Diagnostic Techniques

Structural level

what genetic variants is described when they have problems or mutations in the chromosomal segments or when the structure itself of the chromosome has the mutation.

gain or loss of chromosome segments & rearrangements of the segments of the chromosome

What happens in the structural level? There is what?

molecular level

this genetic variant happens when there is a problem with the nucleotide itself and also the amino acid sequence.

deletion / insertion

an event in the molecular level in which there is a removal or addition of base pairs in the amino acid sequence.

nucleotide repeats

an event in the molecular level in which there is repetition of base pairs in the amino acid.

point mutations

an event in the molecular level in which it involves RFLP or restriction fragment length polymorphism & SNP or single nucleotide polymorphism and is used to detect the base pair differences / point mutations.

silent mutation

this type of point mutation means that there is mutation of base pair in the DNA & RNA but the amino acid is still the same thus the protein is still the same.

Missense mutation

a type of point mutation in w/c there's differences in DNA and mRNA resulting to a different protein (Arg).

nonsense mutation

a type of point mutation in which it affects both DNA and mRNA resulting to a stop codon (meaning no protein is produced)

splice site mutation

this means that there's a damage in splicing on the introns or a change that results in altered RNA sequence.

splicing

meaning, it removes excess introns & stitches introns together with exons

exons

these are the coding portion of the nucleic acid sequence.

introns

these are the noncoding portion of nucleic acid sequence.

base substitution mutation

a mutation in w/c there's a difference in nitrogenous base resulting to one different protein

base deletion mutation

a mutation in w/c there's a deletion or addition of nitrogenous base. More than 1 amino acid is affected leading to either deletion or addition of the nitrogenous base.

Minisatellites or VNTR (Variable Number Tandem Repeats)

this is a type of tandem repeats based on 8-80 base pair repeats.

Microsatellites or STR (Short Tandem Repeats)

a type of tandem repeats based on 2-7 base pair repeats

single nucleotide polymorphism (SNP)

this occurs when there's a single nucleotide different from the reference genetic sequence.

deletion / insertion

this occurs when there is loss of nucleotide

polymorphism

it's the difference in a DNA sequence with atleast 2 alternate forms make it different from each other. this is present in the general population w/ a frequency of > 1%

germ-line mutation

A type of mutation in w/c it is hereditary and can be passed down thru family.

somatic mutation

a type of mutation in w/c it cannot be inherited/

Huntington disease

trinucleotide repeat: CAG

myotonic dystrophy

trinucleotide repeat: CTG

fragile X syndrome

trinucleotide repeat: CGG

spinobular muscular atrophy

trinucleotide repeat: also CAG

restriction endonucleases

a nucleic acid modifying enzyme that cleave double strand DNA

DNA polymerase

a nucleic acid modifying enzyme that synthesizes DNA

DNA ligase function

a nucleic acid modifying enzyme that joins DNA strands together.

kinases

a nucleic acid modifying enzyme that's the phosphorylation of the 5' terminus of the DNA molecule

phosphatase

a nucleic acid modifying enzyme that's the dephosphorylates of the 5' terminus of the DNA

ribonuclease / deoxyribonuclease

a nucleic acid modifying enzyme that aids in the digestion of the DNA

amplification techniques

this involves the PCR and its modifications, and the LAMP

blotting technique

a molecular diagnostic technique that includes SB, NB, WB. RFLP etc.

hybridization technique

a molecular diagnostic technique that includes FISH and microassays

restriction endonucleases

a component in molecular diagnostic technique that will serve as the molecular scissors because it cleave the DNA at a specific sequence.

vectors

a component in molecular diagnostic techniques that serves as a carrier DNA molecule to which the fragment of the DNA of interest is attached.

probes

a component in molecular diagnostic technique that is the pieces of DNA or RNA labelled by various techniques to detect a complementary sequence.

taq polymerase

it's the enzyme for replicating in the PCR

reverse transcription PCR (R

this PCR modification has a starting point of RNA material & an end product of cDNA

nested PCR

a PCR modification that involves 2 sets of primers- outer & inner

multiplex PCR

a PCR modification that uses more than 1 pair of primers.

loop-mediated isothermal amplification (LAMP)

a PCR modification that uses 6 different primers to recognized 8 regions of the target genes.

southern blot

this blotting technique detects DNA

northern blot

this blotting technique detects RNA

western blot

this blotting technique detects proteins

Edwin Southern (1975)

who discovered southern blotting? when?

James Alwine & George Stark (1979)

who developed the northern blotting? when?

variable number tandem repeats (VNTR)

DNA variations that results to RFLP in w/c there is a short sequences of DNA that are repeated multiple times

single nucleotides polymorphism (SNP)

DNA variations that results to RFLP that usually occurs in intervening sequence.

Mutations

DNA variations that results to RFLP that can lead to polymorphism or fragmented cleavage.

hybridization

there's a formation of a duplex between 2 complementary sequences

Fluorescent In Situ Hybridization (FISH)

this type of hybridization is very sensitive

centromeric (satellite) probes

a type of FISH probe in w/c/ fluorescence is in the centromere (center) part of the chromosome

locus specific (telomere) probes

a type of FISH probe in w/c the fluorescence is in the telomere (hands) part of the chromosome

whole chromosome painting probe

a type of FISH probe in w/c almost all parts of chromosomes have fluorescence.

microarray technique

it utilizes next-generation sequencing by use of DNA chip

genotyping

microarray technique in w/c the single-stranded DNA genome is added

gene expression

a microarray technique in which cDNA is added

Rochalimaea henselae

causative agent of bacillary angiomatosis

Ehrlichia chaffeensis

causative agent of human ehrlichiosis

Zika virus

causative agent of zika fever endemic

Influenza A subtype H1N1

caused the 2009 flu pandemic