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2020 MHS Genetics Unit2 > Lab techniques > Flashcards

Flashcards in Lab techniques Deck (25)
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1

detection of VNTR

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

2

detection of a mutation that dirupts RFLPs

southern blot- not sure why

3

detection of an infection specific antigen

ELISA

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

4

analysis/comparison of mutliple genes expression in biological samples

microarray

5

detects a mutation that disrupts specific restriction sites (SSA)

RFLP

6

what are the techniques for identification of specific sequences?

  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

7

DNA fingerprinting 

VNTR

8

Northern blot for RNA and southern blot for DNA

hybeidization

9

used to localize specfici sequences on chromosomes

FISH

10

most ocmmonly used to amplify a known genomic sequence

PCR- used in cloning and expression.

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

11

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

geneomic DNA

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

12

make modification in the exisiting sequence

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

13

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

FISH

  1. CML
  2. CLL

14

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

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

15

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

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

16

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

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

17

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

recombinant antigen 

18

injecting a DNA sequence to ellicit an immune response

DNA vaccine

19

therapy used to replace enzymes 

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

20

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

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

21

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

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

22

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

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

23

what can western blot do?

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 

24

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

SDS-PAGE + mass spectrometery (NOT WESTERN BLOT)

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

25

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