Lecture 4

Question 1

Karyotyping

Answer 1

Chromosome imaging

Question 2

Composition of a Chromosome

Answer 2

Centromere, p and q arm

Question 3

Chromatid

Answer 3

Chromosome during replication or meiosis

Question 4

First step for karyotyping

Answer 4

Using mitotic inhibitor

Question 5

Second step for karyotyping

Answer 5

Lymphocytes are harvested and treated with hypotonic solution

Question 6

Third step for karyotyping

Answer 6

Chromosomes are fixed and dropped onto glass

Question 7

Fourth step for karyotyping

Answer 7

Chromosomes are stained and banding patterns is produced

Question 8

Fourth step for karyotyping

Answer 8

Chromosomes are stained and banding patterns is produced

Question 9

Types of chromsomes’morphology

Answer 9

Metacentric, Submetacentric and acrocentric

Question 10

Tiny DNA of acrocentric chromosome

Answer 10

Satellite stalks

Question 11

Incorrect number of chromosomes

Answer 11

Aneuploidy

Question 12

One reason for reciprocal translocation

Answer 12

Two non homologous chromosomes have double strand breaks + NHEJ

Question 13

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

Answer 13

Philadelphia chromosome (22)

Question 14

FISH acronym meaning

Answer 14

Fluorescence In Situ Hybridization

Question 15

For what metaphase FISH is used

Answer 15

Micro-deletion, micro-insertion, structural rearrangement

Question 16

Name of technique used to detect DNA

Answer 16

Southern blot

Question 17

Name of technique used to detect RNA

Answer 17

Northern blot

Question 18

Name of technique used to detect proteins

Answer 18

Western blot

Question 19

What does restriction enzyme do?

Answer 19

Cut DNA into smaller fragments

Question 20

Three types of restriction enzymes

Answer 20

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

Question 21

How does Southern Blot + Restriction enzymes work?

Answer 21

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

Question 22

What does PCR stands for?

Answer 22

Polymerase Chain Reaction

Question 23

Steps of PCR

Answer 23

Denaturation of DNA, primer annealing, DNA synthesis

Question 24

Promoter

Answer 24

Sequence of a gene where RNA polymerase fixes before starting transcription

Question 25

What molecule is used to sequence RNA?

Answer 25

Using cDNA

Question 26

What does IP stand for

Answer 26

Immunoprecipitation

Question 27

What is immunoprecipitation

Answer 27

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

Question 28

What is Co-IP?

Answer 28

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

Question 29

What does CGH (microarray) stand for?

Answer 29

Comparative Genomic Hybridization

Question 30

Limitations of array technology

Answer 30

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

Question 31

Steps of Sanger sequencing

Answer 31

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

Question 32

What is NGS?

Answer 32

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

Question 33

Sequence detection technologies

Answer 33

- Sanger sequencing - 454 pyrosequencing - Illumina (Solexa) Sequencing - SOLiD sequencing - Ion Torrent sequencing

Question 34

What is Bridge PCR?

Answer 34

72/94

Question 35

Construction of recombinant DNA molecules in vitro

Answer 35

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

Question 36

How do we select only the bacteria with DNA inserted?

Answer 36

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

Question 37

What is a library

Answer 37

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

Question 38

What contain a cDNA library

Answer 38

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

Question 39

What is the main way to create a library?

Answer 39

Use of Bacterial Artificial Chromosome

Question 40

What is BAC?

Answer 40

Bacterial Artificial Chromosome

Question 41

What are the characteristics of a BAC?

Answer 41

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

Question 42

What are the two genetic approaches in model systems?

Answer 42

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

Question 43

Approach to do Forward genetics

Answer 43

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