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Principle of genetics > Lecture 4 > Flashcards

Lecture 4 Flashcards

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Biology 101 flashcards
1
Q

Karyotyping

A

Chromosome imaging

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2
Q

Composition of a Chromosome

A

Centromere, p and q arm

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3
Q

Chromatid

A

Chromosome during replication or meiosis

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4
Q

First step for karyotyping

A

Using mitotic inhibitor

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5
Q

Second step for karyotyping

A

Lymphocytes are harvested and treated with hypotonic solution

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6
Q

Third step for karyotyping

A

Chromosomes are fixed and dropped onto glass

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7
Q

Fourth step for karyotyping

A

Chromosomes are stained and banding patterns is produced

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8
Q

Fourth step for karyotyping

A

Chromosomes are stained and banding patterns is produced

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9
Q

Types of chromsomes’morphology

A

Metacentric, Submetacentric and acrocentric

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10
Q

Tiny DNA of acrocentric chromosome

A

Satellite stalks

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11
Q

Incorrect number of chromosomes

A

Aneuploidy

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12
Q

One reason for reciprocal translocation

A

Two non homologous chromosomes have double strand breaks + NHEJ

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13
Q

Example of a reciprocal translocation of chromosomes (leading to Chronic Myeloid Leukemia)

A

Philadelphia chromosome (22)

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14
Q

FISH acronym meaning

A

Fluorescence In Situ Hybridization

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15
Q

For what metaphase FISH is used

A

Micro-deletion, micro-insertion, structural rearrangement

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16
Q

Name of technique used to detect DNA

A

Southern blot

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17
Q

Name of technique used to detect RNA

A

Northern blot

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18
Q

Name of technique used to detect proteins

A

Western blot

19
Q

What does restriction enzyme do?

A

Cut DNA into smaller fragments

20
Q

Three types of restriction enzymes

A

Blunt ends, 5’ sticky ends, 3’ sticky ends

21
Q

How does Southern Blot + Restriction enzymes work?

A

Restriction enzyme cut after a specific sequence. If not present, not cut will be there and southern blot will detect longer dna

22
Q

What does PCR stands for?

A

Polymerase Chain Reaction

23
Q

Steps of PCR

A

Denaturation of DNA, primer annealing, DNA synthesis

24
Q

Promoter

A

Sequence of a gene where RNA polymerase fixes before starting transcription

25
Q

What molecule is used to sequence RNA?

A

Using cDNA

26
Q

What does IP stand for

A

Immunoprecipitation

27
Q

What is immunoprecipitation

A

Technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein

28
Q

What is Co-IP?

A

Selects an antibody that targets a known protein that is believed to be a member of a larger complex of proteins

29
Q

What does CGH (microarray) stand for?

A

Comparative Genomic Hybridization

30
Q

Limitations of array technology

A
  1. Can’t detect balanced chromosomal rearrangements (reciprocal translocations, inversions, polyploidy)
  2. Do not cover highly repeat-rich sequences
31
Q

Steps of Sanger sequencing

A
  1. PCR
  2. Chop DNA
  3. DNA with nucleotides and (small amount) dideoxynucleotide (stop DNA chain elongation)
  4. Reading of dye-labelled fluorescent tags on ddNucl
32
Q

What is NGS?

A

Next Generation Sequencing: Simultaneous sequencing of hundreds of millions of short sequences

33
Q

Sequence detection technologies

A
  • Sanger sequencing
  • 454 pyrosequencing
  • Illumina (Solexa) Sequencing
  • SOLiD sequencing
  • Ion Torrent sequencing
34
Q

What is Bridge PCR?

A

72/94

35
Q

Construction of recombinant DNA molecules in vitro

A
  1. Cleave vector and insert DNAs with compatibles sticky ends
  2. Ligation of DNA pieces
  3. Bacterial transformation => Bacteria are mixed with the recombinant plasmid and will inherit this recombinant DNA
36
Q

How do we select only the bacteria with DNA inserted?

A

DNA inserted has also gene for penicilin resistance => Only these are alive

37
Q

What is a library

A

A library is a way to store genetic information or a way to do genetic screening

38
Q

What contain a cDNA library

A

It contains the expressed (or transcribed) genes of a given tissue

39
Q

What is the main way to create a library?

A

Use of Bacterial Artificial Chromosome

40
Q

What is BAC?

A

Bacterial Artificial Chromosome

41
Q

What are the characteristics of a BAC?

A
  • Vector of choice for current genomic library construction and screening
  • Holds 100 to 200kb
  • Stably propagated in E. coli
42
Q

What are the two genetic approaches in model systems?

A
  1. Forward genetics: Which genes are responsible for a given phenotype
  2. Reverse genetics: What change in the phenotype is observed when a gene is altered
43
Q

Approach to do Forward genetics

A
  1. Create genetic diversity
  2. Select the cells/organisms with the interesting phenotypes
  3. Identify genes involved

Principle of genetics (9 decks)

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