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Molbio. > PV92 > Flashcards

PV92 Flashcards

1
Q

biorad manual

PCR uses heat-resistant Taq polymerase from?

A

Thermus aquaticus

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2
Q

biorad manual

volume of saline to be transfer in the micro test tube.

how long will u rinse ur mouth using saline?

A

1 ml

30 seconds

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3
Q

sample of choice

A
  • hair follicles
  • oral epithelial cells (cheek cells)
  • drop of blood
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4
Q

centrifuge time for saline

what u should see after centrifugation?

A

2 minutes

match-head sized pellet of whitish cells at the BOTTOM of the tube.

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5
Q

biorad manual

temp where it loosens connective tissue and inactivates DNAses

hint: temp of water bath

A

56C

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6
Q

biorad manual

ruptures cell membranes and denatures protein

this is the temp of?

A

100C

dry bath

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7
Q

discussion

PV92 PCR is to detect — which composed of — nucleotides

A

to detect alu transposon
composed of 300 nucleotides

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8
Q

discussion

marker of human migration

what is the purpose?

A

Alu I restriction enzyme

relatedness among individuals

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9
Q

discussion

locus/region of DNA is called?

A

PV92

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10
Q

discussion

GIVE THE GENOTYPE AND SIZE OF PCR PRODUCTS
- (+/+)
- (-/-)
- (+/-)

A
  • (+/+) = homozygous 941 bp
  • (-/-) = homozygous 641 bp
  • (+/-) = heterozygous 941 and 641 bp
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11
Q

discussion

what is the purpose of instagene matrix?

A

to lyse cheek cells

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12
Q

discussion

after adding instagene matrix, what’s the next step?

A

dry bath 100C for 5 mins

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13
Q

discussion

contents of master mix (6)

A
  • DNA pol (Taq polymerase)
  • magnesium ion (enzyme cofactor)
  • PCR primer
  • salt buffer
  • template
  • nucleotides (dATP, dCTP, dGTP, dTTP)
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14
Q

discussion

why need control?

A

to check the reagent

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15
Q

discussion

regulates temperature of the sample in thermal cycler

A

aluminum blocks

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16
Q

discussion

used for staining of agarose gel

A

orange G stain

17
Q

discussion

how do you prepare gel casting tray?

A
  1. comb to create wells
  2. buffer
  3. set the volts
18
Q

discussion

how do you report?
- homozygous
- homozygous (-)
- heterozygous

A

can be positive/negative
- homozygous = mm
- homozygous (-) = mn
- heterozygous = nn

19
Q

discussion

enumerate the steps in genomic DNA protocol

separate in each phase

A

SAMPLE PREP
1. sample (if oral, use saline mouth wash)
2. cell lysis
3. centrifuge to separate the cells
3. add instagene matrix to lyse check cells
4. dry bath 100C for 5 mins
5. water bath 56C for 10 mins

REAGENT PREP
1. master mix prep

PCR AMPLIFICATION/ANALYSIS
1. denaturation
2. annealing
3. extension
4. gel electrophoresis
5. stain the gel
6. visualize the gel under UV

should have a band that indicates PCR was successful.

20
Q

discussion

enumerate the steps in gel electrophoresis

what’s the complete name of the electrophoresis we used?

A
  1. prepare agarose
  2. prepare gel cating tray
  3. electrophoresis prep
  4. electrophoresis loading
  5. staining of agarose gel

agarose gel electrophoresis

21
Q

discussion

principle stating that the genetic variation in a population will remain constant from one generation to the nex in the absence of distrurbing factors.

A

Hardy-Weinberg Equilibrium

p^2 + 2pq + q^2 = 1

22
Q

discussion

how to calculate genotypic frequencies?

A

number of category / total number

23
Q

discussion

negatively charged, microscopic beads that chelate or grab, metal ions out of solution. traps metal ions, such as Mg2+ which are required as catalysts or cofactors in enzymatic reactions

but what is the function?

A

instagene matrix

binds cellular magnesium ions

24
Q

biorad manual

PV92 is used in?

A
  • population genetics
  • paternity analysis
  • forensics
25
Q

lab manual

temp for each step of PCR

A
  1. denaturation = 94-98C
  2. annealing = 50-65C
  3. extension = 72-75C

remember! should repeat 20-40 cycles

26
Q

biorad manual

PV92 locus is on chromosome?

A

chromosome 16

27
Q

biorad manual

approx. — Alu copies per haploid genome, representing about — of the genome

A

500,000 Alu copies per haploid, representing about 5% of the genom

28
Q

biorad manual

genotypes

A
  • +/-
  • -/-
  • +/-
29
Q

biorad manual

determionation and analysis of alu frequency in a population

enumerate step-by-step

A
  1. amplify Alu insert from represenative sample population
  2. calculate the expected allelic and genotypic frequencies
  3. perform chi-square test

Molbio. (6 decks)

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