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Biology Exam > Molecular Methods > Flashcards

Molecular Methods Flashcards

1
Q

Usage of methods of molecular biology

A 
Discovery of new genes & proteins
Study of gene regulation & protein function
Evolution study
Diagnosis of pathogens
Diagnosis of genetic diseases
Production of medicaments
Food industry
Gene engineering
Forensic medicine
Criminalistics
Identification of animals 
Pedigree tests
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2
Q

Usage of methods of molecular biology - evolution study

A

Phylogenetic maps

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3
Q

Usage of methods of molecular biology - diagnosis of genetic diseases

A

DNA mutations

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4
Q

Usage of methods of molecular biology - production of medicaments

A

Vaccine, insulin

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5
Q

Usage of methods of molecular biology - food industry

A

Identification of ingredients

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6
Q

Usage of methods of molecular biology - gene engineering

A

production of GMO

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7
Q

Usage of methods of molecular biology - forensic medicine

A

paternity & maternity tests

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8
Q

Usage of methods of molecular biology- criminalistics

A

Identification of persons

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9
Q

Usage of methods of molecular biology - identification of animals

A

Illegal tranport

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10
Q

DNA Isolation material

A

Virus,bacteria, parasites, plants, animals
Body fluids
Tissues, hairs, feather
Fresh/frozen/in parafin

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11
Q

Cloning

A

Formation of new copies (clone)

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12
Q

Cloning types

A

Molecular
Embryonal
Transnuclear

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13
Q

Cloning - molecular

A

Copying DNA by PCR or copying in bacteria

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14
Q

Cloning - embryonal

A

Division of embryonal cells that results in identical twins

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15
Q

Cloning - transnuclear

A

Transfer of nucleus from somatic cell into sex cell results in forming the copy of individuum

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16
Q

PCR PRinciple

A

Forming of identical copy of DNA

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17
Q

PCR Prepare Mixture

A 
Isolated DNA
Synthetic primers
Nucleotides
Taq polymerase 
Buffer with Mg2+ ions
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18
Q

Gel electrophoresis principle

A

Separation of DNA fragments according to their size

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19
Q

Gel electrophoresis procedure

A

Prepare agarose gel (add ethidium bromide)
Stain sample (w/ bromphenol blue)
Lay sample into gel, set electrophoresis
Visualize DNA fragments under UV transluminator

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20
Q

DNA Cloning in bacteria

A

Small circular DNA with:
Replication origin
Place for restriction endonuclease
Gene for some specific characteristic (antibiotic resistance)

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21
Q

RFLP procedure

A

Cleavage of DNA by restriction endonuclease

Analysis of DNA fragments on agarose gel

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22
Q

Usage of RFLP

A

Paternity & maternity tests
Identification of individuum
Identification of bird sex
Studyy of polymorphism & relationship

23
Q

Southern blotting principle

A

Transfer of ssDNA from gel into nylon membrane by equipment “vacuum blotter” under vacuum

24
Q

Northern blotting

A

Analysis of RNA

25
Western blotting
Analysis of proteins
26
Hybridisation principle
Specific DNA probe is bound to ssDNA based on complementarity
27
DNA probe
ssDNA | Marked radiactive or non-radioactive
28
Hybridisation procedure
Nylon membrane with blotted DNA is closed into plastic bag together with marked DNA probe Hybridisation of probe Removing of non-bound probe from nylon membrane Detection - autoradiography or luminescence
29
DNA Sequencing principle
Setting precise order of nucleotides in DNA in equipment "sequenator"
30
Methods of sequencing
Maxam-Gilbert sequencing | Sanger sequencing
31
Sanger sequencing
Faster, simpler, no use of toxic substances | Determines the nucleotide order of a given DNA fragment
32
Sanger sequencing method
Uses same principle as PCR, when template DNA is amplified by help of primers & polymerase During sequencing, just 1 primer is used & that is why just 1 DNA strand is synthesised in 1 direction Sequencing reaction takes place in 4 tubes (4 reactions) Result is mixture of DNA fragments of different size starting with primer & ending with certain dideoxynucleotide DNA fragments can be separated based on its size by gel electrophoresis & set nucleotide order
33
Sanger sequencing ingredients
``` Template DNA Primer DNA polymerase Standard deoxynucleotides 1 of the 4 dideoxynucleotide triphosphates ```
34
Dideoxynucleotide
Modified nucleotide Lacking a 3'-OH group required for the formation of phosphodiester bond between 2 nucleotides. thus terminating DNA strand extension & resulting in DNA fragments of varying lengths
35
Automatic sequence
Variant of enzymatic DNA sequencing Dideoxynucleotides are marked with different fluorescent colors (4 diff. colors) DNA synthesis takes place in 1 reaction Capilar electrophoresis is used to separate DNA fragments based on their size, fluorescent color is detected by detector at the end of capilar & thus set the nucleotide order
36
Application of DNA sequencing
``` Research Medicine Phylogenesis Anthropology Criminalistic Forensic science Agriculture ```
37
DNA sequencing - research
Sequencing of whole genome or just part of genome (study of evolution)
38
DNA sequencing - medicine
Diagnosis of genetic diseases & predispositions (cancer, cardiovascular diseases), gene therapy
39
DNA sequencing - phylogenesis
Study of evolution
40
DNA sequencing - Anthropology
DNA comparison to find out migration of human king (mitochondrial DNA, Y-chromosomal DNA)
41
DNA sequencing - sequencing
Using of repetitive sequences of DNA (microsatellites, minisatellites)
42
DNA sequencing - forensic medicine
Paternity test
43
DNA sequencing - agriculture
GMO
44
GMO
Organism whose genetic material was altered using genetic engineering techniques (recombinant DNA technology)
45
What is DNA microchip?
Small glass plate with 1000s of short synthetic ssDNA (for diff. genes)sequences on the surface
46
What is a DNA microchip used for?
Research tool used to identify mutations in genes that can cause disease Researchers will produce chips to help assess individual risks for developing different cancers, heart diseases, diabetes, etc.
47
How does a DNA microchip work?
Obtain DNA from patient's blood & control DNA without mutation in monitored genes Denature DNA, cut into small fragments &label them by fluorescent dye Both sets of labeled DNA are inserted into chip & allowed to hybridise to the synthetic monitored genes on the chip
48
Do twins have identical DNA?
Identical twins (monozygotic) Formed when 1 fertilised egg splits Identical DNA Different phenotype - result of interaction between genes & environment in the uterus DNA test can't determine the difference while fingerprint can
49
Chimera in greek myhtology
Monstrous creature that combined the parts of goat, lion, serpent
50
Chimera in genetics
Animal that has 2 or more different populations of genetically distinct cells that originated in different zygotes
51
Human chimeras - blood chimeras
People with more than 1 blood type
52
Human chimeras - identical twins
Embryos share blood supply in placenta, allowing blood stem cells to pass from one and settle in bone marrow of other
53
Human chimeras - non-identical twins
Blood cells come from twin that died in early gestation

Biology Exam (14 decks)

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