Lab techniques

Question 1

detection of VNTR

Answer 1

PCR

VTNR- variable tandem number repeat. these can be detected by size discrepancey.

1. RFLP cuts the flanking regions.
2. more number of repeats -> travels shorter distance.
3. highly specific per person

Question 2

detection of a mutation that dirupts RFLPs

Answer 2

southern blot- not sure why

Question 3

detection of an infection specific antigen

Answer 3

ELISA

1. Antigens are protein, most of the time
2. fastest

Question 4

analysis/comparison of mutliple genes expression in biological samples

Answer 4

microarray

Question 5

detects a mutation that disrupts specific restriction sites (SSA)

Answer 5

RFLP

Question 6

what are the techniques for identification of specific sequences?

Answer 6

1. SANGER sequencing
2. NGS
3. RFLP- used to detect a presence or absence of restriction site(s) in the sequence of interest (known sequence)
4. VNTR- identifies the variable number tandem random repeats
   
   1. done by cloning enzymes or PCR
   2. DNA fingerprinting

Question 7

DNA fingerprinting

Answer 7

VNTR

Question 8

Northern blot for RNA and southern blot for DNA

Answer 8

hybeidization

Question 9

used to localize specfici sequences on chromosomes

Answer 9

FISH

Question 10

most ocmmonly used to amplify a known genomic sequence

Answer 10

PCR- used in cloning and expression.

1. eukaryotic gene cannot be used for protein expression because of introns

Question 11

used to identify SNPs and other mutation or alterations in nucleic acids

Answer 11

geneomic DNA

1. can be amplified by PCR if the sequence of interest is known -most common technique

Question 12

make modification in the exisiting sequence

Answer 12

chemical synthesis is primarily to make modific ation in the existing sequence to optimize gene expression in heterological system.

Question 13

used to detect chromosomal aberrations and to identify a location of specific sequences on chromosomes

Answer 13

FISH

1. CML
2. CLL

Question 14

used to determine the pharmakogenetics of an indivudal for proper medicine dosage

Answer 14

microarrays

1. can be used to
   
   1. compare the expression pattern of cancerous and normal cells
   2. analyze drug metabolism
      
      1. different CYP450 expressions
   3. differences in expression patterns of gene expression of the same type of cancer from different patients, which can assist with choosing the most efficient therapy

Question 15

May be used to assist physician to probable reaction to a cancer treatment.

Answer 15

microarrays

1. can be used to
   
   1. compare the expression pattern of cancerous and normal cells
   2. analyze drug metabolism
      
      1. different CYP450 expressions
   3. differences in expression patterns of gene expression of the same type of cancer from different patients, which can assist with choosing the most efficient therapy

Question 16

used to compare the expression levels of all genes or a set of specific set of genes in different cell types

Answer 16

microarrays

1. can be used to
   
   1. compare the expression pattern of cancerous and normal cells
   2. analyze drug metabolism
      
      1. different CYP450 expressions
   3. differences in expression patterns of gene expression of the same type of cancer from different patients, which can assist with choosing the most efficient therapy

Question 17

antigenic proteins expressed outside of the host and used in a vaccine

Answer 17

recombinant antigen

Question 18

injecting a DNA sequence to ellicit an immune response

Answer 18

DNA vaccine

Question 19

therapy used to replace enzymes

Answer 19

recombinant proteins used in enzyme replacement therapy or as signaling molecules; the choice of expression system depends on the prottein complexity and post-translational modifications

Question 20

analysis of troponin or HIV in a biological sample yeilds results with in seconds-minutes. What what the test?

Answer 20

ELISA

1. quantitative
2. easy
3. fast
4. analyze large number of samples for detection of
   
   1. suspected pathogens: bacterial or viral
   2. disease-related protein markers
      
      1. troponin
      2. Ab
      3. tumor markers

Question 21

quantitative fast and detects proteins in biological samples. Can this test discriminate between mutant and normal?

Answer 21

ELISA

1. quantitative
2. easy
3. fast
4. analyze largenumber of samples for detection of
   
   1. suspected pathogens: bacterial or viral
   2. disease-related protein markers
      
      1. troponin
      2. Ab
      3. tumor markers
5. this test cannot differentiate between normal and mutant

Question 22

A post-translational modification leads to a disease. What is the best methodology to observe the normal vs muant?

Answer 22

western blot

1. used to
   
   1. identify a truncated mutatnt protein
   2. different from ELISA
      
      1. ELISA will detect the normal and mutant with out discriminating
      2. western blot will differentiate between the mutant and normal
   3. the mutant and normal will migrate to different locations in the gel

Question 23

what can western blot do?

Answer 23

western blot

1. used to
   
   1. identify a truncated mutatnt protein
   2. different from ELISA
      
      1. ELISA will detect the normal and mutant with out discriminating
      2. western blot will differentiate between the mutant and normal
2. using SDS-PAGE to run the samples, tranfer to nitrocellulose, and visualization using the antibodies

Question 24

I desire to separate my protein samples and identify them. What method is the best option?

Answer 24

SDS-PAGE + mass spectrometery (NOT WESTERN BLOT)

1. remeber, WB uses nitrocellulose to visualize the sample, which cannot be used for mass spec

Question 25

A mother, 6 months pregnant, wants to know if her has a certain disease gene. What can be done with little invasivness?

Answer 25