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Genetics > Tools of Molecular Genetics and Cytogenetics > Flashcards

Tools of Molecular Genetics and Cytogenetics Flashcards

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1
Q

What two fundamental obstacles do geneticists face?

A

Obtaining enough DNA or RNA sequence to analyze

Purification of sequence of interest

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2
Q

What is a process of isolating a particular DNA sequence and creating multiple copies in vivo?

A

Molecular cloning

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3
Q

What are 3 elements required for molecular cloning?

A

Origin of replication
Selectable marker (sort which are carrying DNA)
Other features are desired

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4
Q

What are the four steps of molecular cloning?

A

DNA fragmentation w/ restriction endonucleases
Ligation of DNA fragments to a vector
Transfection/ transformation
Screening/ selection

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5
Q 
These are all ways to what?
•PCR
•Restriction enzyme digestion
•DNA sonication and fractionation 
•Chemically synthesized oligionucleotides
A

Isolate DNA fragment of interest

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6
Q

How does selection usually occur?

A

Through antibiotic resistance markers (allow only cells which have the vector to survive)

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7
Q

WHat is a collection of cloned cDNA fragments inserted into host cells?

A

cDNA library

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8
Q

What is a cDNA library produced from?

A

Fully transcribed mRNA (can be expressed in prokaryotic organisms)

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9
Q

What does gel electrophoresis do?

A

Separate by size

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10
Q

What is southern blotting used for?

A

Check for the presence of a DNA sequence in a DNA sample

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11
Q

What does Southern blotting involve?

A

Agarose electrophoresis
Transfer methods
Probe hybridization

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12
Q

What is improper chromosome number?

A

Aneuploidy

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13
Q

What is it when homologues fail to separate?

A

Nondisjunction

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14
Q

What is a monosomic gamete?

A

Has no chromosomes in cell

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15
Q

What is in a trisomic gamete?

A

2 chromosomes (will eventually create a trisomy)

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16
Q

What is the term for when one chromosome is left out of new nucleus due to lag?

A

Anaphase lag

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17
Q

What is the term for too many chromosomes?

A

Polysomy

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18
Q

What is the term for exchange of portions of non-homologous chromsomes?

A

Translocation

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19
Q

What is removal and reverse attachment of a portion of a chromosome?

A

Inversion

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20
Q

Are autosomal monosomies compatable with life?

A

No

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21
Q

Are autosomal polysomies compatible with life?

A

Yes (but typically have a shorter life span)

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22
Q

Are sex chromosome monosomies and trisomies compatible with life?

A

Yes

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23
Q

How much of human DNA is junk?

A

95%

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24
Q

Where do most mistake in chromosome structure occur?

A

Crossing over

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25
Q

What are the four common types of chromosome damage?

A

Translocation
Inversion
Deletion
Duplication

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26
Q

What is Trisomy 21?

A

Down Syndrome

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27
Q

What is the leading cause of mental retardation?

A

Down Syndrome

28
Q

What are the three autosomal trisomies that are survivable?

A 
Down syndrome (21)
Edwards syndrome (18) 
Trisomy 13- Patau syndrome
29
Q

What is the rare form of Down Syndrome?

A

Translocation on long arm of chromosome 21 to another chromosome

30
Q

What syndrome is Trisomy 18?

A

Edwards syndrome (less common and more severe than Trisomy 21) only live a few weeks

31
Q

What is Trisomy 13?

A

Patau syndrome

32
Q

What syndrome is associated with a deletion of short arm of chromosome 5

A

Cri du Chat syndrome

33
Q

What does a child with cri du chat look like?

A

Severe retardation
Round face
Heart anomalies
Cry like cat

34
Q

What is monosomy X?

A

Turner syndrome

35
Q

What are the manifestations of monosomy X?

A 
Short stature
Webbed neck
Fibrous ovaries
Sterility
Amenorrhea
Wide chest
Heart defects
36
Q

What occurs with multiple X females?

A

XXX
Normal appearance
Menstrual abnormalities

37
Q

What is the only different in males with XYY?

A

Taller than average

38
Q

Big defining feature of Turner syndrome?

A

Webbed neck

39
Q

What is the best probe to use in DNA analysis to look for a specific mutation?

A

Synthetic oligonucleotide

40
Q

What is an ASO probe?

A

Allele-specific oligonucleotide

41
Q

What is an ASO used for?

A

To see changes too small for traditional Southern blot analysis
Can tell apart normal homozygous, mutant homozygous, heterozygous

42
Q

What technique is used to analyze RNA?

A

Nothern/ RNA Blotting

43
Q

Is there endonuclease cleavage in Nothern Blotting?

A

No

44
Q

What is used to amplify single or few copies of a piece of DNA by several order of magnitude?

A

PCR (polymerase chain reaction)

45
Q

What does PCR involve?

A

Thermal cycling and enzymatic replication

46
Q

PCR requires ______ complementary to target region and DNA polymerase

A

primers

47
Q

What polymerase survives thermal cycling and is used in PCR?

A

Taq polymerase

48
Q

In PCR what does heat do?

A

Separates DNA strands to create templates

49
Q

What happens in cooler temperatures in PCR?

A

DNA synthesis

50
Q

What is the study of chromosomes, their structure, and their inheritance?

A

cytogenetics

51
Q

What does a cDNA library not have information about?

A

Enhancers
Introns
Other regulatory elements

52
Q

What enzyme creates mature mRNA from cDNA?

A

Reverse transcriptase

53
Q

In which type of blotting is DNA gel placed in alkaline solution to denature the ds-DNA?

A

Southern Blotting

54
Q

In Southern Blotting, what causes DNA to move from gel onto membrane

A

Capillary action with appropriate buffer

55
Q

What binds the DNA to the membrane in Southern Blotting?

A

Ion exchange

56
Q

In Southern Blotting, what indicates that the fragment contains DNA sequence complementary to the probe?

A

Hybridization of the probe to a DNA fragment on the filter membrane

57
Q

Why is it difficult to identify mutations in southern blotting?

A

Only those that affect the size of the fragment are seen

58
Q

What type of blotting doesn’t involve endonuclease cleavage?

A

Northern Blotting

59
Q

What is Klinefelter syndrome?

A

Where a male has an extra X- causes feminization

60
Q

What is the order of vectors from smallest to largest

A

Plasmids –> phages –> cosmids –> YAC

61
Q

What type of vector is grown in yeast?

A

YAC

62
Q

What type of vector is when a virus infects bacteria?

A

Phage

63
Q

What type of vector is a larger plasmid?

A

Cosmid

64
Q

what type of vector must go through transfection?

A

YAC

65
Q

Southern Blotting only identifies mutations that affect the ____ of the DNA fragment.

A

Size

Genetics (10 decks)

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