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Biology Exam > Molecular Methods > Flashcards

Molecular Methods Flashcards

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AP® Biology flashcards Biology 101 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
Q

Western blotting

A

Analysis of proteins

26
Q

Hybridisation principle

A

Specific DNA probe is bound to ssDNA based on complementarity

27
Q

DNA probe

A

ssDNA

Marked radiactive or non-radioactive

28
Q

Hybridisation procedure

A

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
Q

DNA Sequencing principle

A

Setting precise order of nucleotides in DNA in equipment “sequenator”

30
Q

Methods of sequencing

A

Maxam-Gilbert sequencing

Sanger sequencing

31
Q

Sanger sequencing

A

Faster, simpler, no use of toxic substances

Determines the nucleotide order of a given DNA fragment

32
Q

Sanger sequencing method

A

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
Q

Sanger sequencing ingredients

A 
Template DNA
Primer
DNA polymerase
Standard deoxynucleotides
1 of the 4 dideoxynucleotide triphosphates
34
Q

Dideoxynucleotide

A

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
Q

Automatic sequence

A

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
Q

Application of DNA sequencing

A 
Research
Medicine
Phylogenesis Anthropology
Criminalistic
Forensic science
Agriculture
37
Q

DNA sequencing - research

A

Sequencing of whole genome or just part of genome (study of evolution)

38
Q

DNA sequencing - medicine

A

Diagnosis of genetic diseases & predispositions (cancer, cardiovascular diseases), gene therapy

39
Q

DNA sequencing - phylogenesis

A

Study of evolution

40
Q

DNA sequencing - Anthropology

A

DNA comparison
to find out migration of human king (mitochondrial
DNA, Y-chromosomal DNA)

41
Q

DNA sequencing - sequencing

A

Using of repetitive sequences of DNA (microsatellites, minisatellites)

42
Q

DNA sequencing - forensic medicine

A

Paternity test

43
Q

DNA sequencing - agriculture

A

GMO

44
Q

GMO

A

Organism whose genetic material was altered using genetic engineering techniques (recombinant DNA technology)

45
Q

What is DNA microchip?

A

Small glass plate with 1000s of short synthetic ssDNA (for diff. genes)sequences on the surface

46
Q

What is a DNA microchip used for?

A

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
Q

How does a DNA microchip work?

A

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
Q

Do twins have identical DNA?

A

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
Q

Chimera in greek myhtology

A

Monstrous creature that combined the parts of goat, lion, serpent

50
Q

Chimera in genetics

A

Animal that has 2 or more different populations of genetically distinct cells that originated in different zygotes

51
Q

Human chimeras - blood chimeras

A

People with more than 1 blood type

52
Q

Human chimeras - identical twins

A

Embryos share blood supply in placenta, allowing blood stem cells to pass from one and settle in bone marrow of other

53
Q

Human chimeras - non-identical twins

A

Blood cells come from twin that died in early gestation

Biology Exam (14 decks)

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