lab 5 Flashcards

(30 cards)

1
Q

what areas did PCR have an impact on

A
  • gene mapping
  • cloning
  • DNA sequencing
  • gene detection
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2
Q

PCR template

A

any form of ds DNA

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3
Q

PCR ingredients

A
  • reaction buffer
  • DNA subunits
  • DNA polymerase
  • 2 DNA primers
  • template strand
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4
Q

what 2 basic processes does PCR make use of

A
  1. complementary DNA strand hybridization
  2. DNA strand synthesis via DNA polymerase
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5
Q

DNA polymerase used

A

Taq polymerase from Thermus aquaticus

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6
Q

steps of PCR

A
  1. denaturation
  2. primer annealing
  3. primer extension
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7
Q

term for PCR machine

A

thermal cycler

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8
Q

temperature/thermal cycling

A

rapid heating and cooling of thermal block in thermal cycler

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9
Q

temp at denaturation step

A

94 degrees Celsius

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10
Q

temp at annealing step

A

60 degrees Celsius

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11
Q

temp at extension step

A

72 degrees Celsius

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12
Q

temperature cycle =

A

denaturation + annealing + extension

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13
Q

PCR generates DNA of a…

A

precise length and sequence

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14
Q

which cycle are fragments of precise length generated

A

third

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15
Q

what are targets of random insertions?

A

introns

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16
Q

what does SINE stand for

A

short repetitive interspersed elements

17
Q

ex. of SINE

18
Q

where does Alu name come from

A

Alu I restriction enzyme recognition site found in this sequence

19
Q

where are we looking for Alu sequence

A

PV92 locus on chromosome 16

20
Q

dimorphic

A

Alu element present in some ind. but not others

21
Q

homozygous Alu presence

22
Q

homozygous Alu absence

23
Q

heterozygous Alu

A

both 641 and 941 bp

24
Q

bioinformatics

A

discipline that integrates mathematical, statistical, and computer tools to collect/process biological data

25
InstaGene matrix
consists of suspension of negatively charged beads which bind divalent cations - need to remove divalent cations because they will assist enzymes in DNA degradation
26
supernatant
contains clean, intact genomic DNA
27
functions of incubation steps
1. breakup of connective tissue 2. inactivates DNases
28
master mix
mixture of nucleotides, buffer, and Taq DNA pol
29
dye that makes DNA primers yellow
tartrazine
30
is manual PCR possible?
yes