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Biotech > Quiz 2 PPTs > Flashcards

Quiz 2 PPTs Flashcards

1
Q

What can go from RNA to DNA?

  • …
  • …
  • …
A

retroviruses;
retroposons;
telomeres

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2
Q

For reverse transcription:

  1. Template: …
  2. primer: …, …, …
A

RNA;

oligo dTs; random hexamers; gene specific

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3
Q

For reverse transcription:

  1. … (aka …)
  2. …
A

RNA-dependent DNA polymerase; Reverse Transcriptase;

nucleotides

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4
Q

RT-PCR: ….

A

reverse transcription-polymerase chain reaction

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5
Q

RT-PCR:

  1. … RNA
  2. … RNA to generate …
A

isolate;

reverse-transcribe; cDNA

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6
Q

RT-PCR:

  1. cDNA becomes … for PCR to … specific sequence
  2. analyze RT-PCR products on …
A

template;
amplify;
agarose gels

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7
Q

RT-PCR:

- analyze RT-PCR products on agarose gels - note that this tells us … if the gene is …, but cannot ….

A

qualitatively;
expressed;
quantitate expression

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8
Q

4 steps of reverse transcription lab:

  1. …
  2. …
  3. …
  4. …
A

prep;
anneal;
extend;
inactivate

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9
Q

RT lab:

+RT is a …; reaction contains …

A

positive control;

all components

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10
Q

RT lab:

  • RT: …, no …
  • RNA: …, no …
A

negative control;
reverse transcriptase;
negative control;
RNA

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11
Q

RT lab:

- RT: Since no RT in reaction, no … can be made. Therefore, any product in PCR is due to …

A

cDNA;

gDNA contamination

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12
Q

RT lab:
- RNA: Since no RNA in reaction, nothing is there to … Therefore, any product in PCR is due to one of the … being …

A

provide a template;
reaction components;
contaminated

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13
Q

expected results for RT PCR:
+RT: …, …
- RT: …, … after pCR
- RNA: …, … after PCR

A

cDNA; product;
no cDNA; no product;
no cDNA; no product

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14
Q

RT lab:

… primers: a pair of primers that anneal to the sequence of the gene of interest and help … that sequence in PCR

A

gene specific;

copy

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15
Q

RT lab:

- Regarding primers, where to start depends on …

A

what it is that you want to do (what you want to copy)

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16
Q

RT lab:
- If you want to amplify a specific region, to make the primers, you need to first … that region and choose primers … that region that best fit the parameters

A

identify;

flanking

17
Q

RT lab:

  • forward primer: …
  • reverse primer: …
A

direct sequence;

reverse and complement

18
Q

RT lab:
PCR primer design:
- … to … nucleotides in length
…-…% GC content, melting temp … - … degrees C

A

18; 24;
40; 60;
45; 65

19
Q

RT lab:
PCR primer design:
- …, …-… Gs or Cs at … end

A

GC clamp;

2-3; 3’

20
Q 
RT lab: 
primer secondary structure: 
- ... 
- ... 
- ... 
also want to avoid ... and ...
A

hairpins;
self dimers;
cross dimers;
runs; repeats

21
Q

For RT-PCR, using … as template

A

single, isolated sequence

22
Q

NanoDrop 2000:
this is the ….
- use …-… microliters of sample
- gives us …

A

spectrophotometer;
1; 2;
concentration

23
Q

RT lab:
NanoDrop 2000:
gives us … and …

A

260/280;

260/230 ratios

24
Q

The aborbance for a molar concentration of a substance with a path length of l cm determined at a …
Its value is obtained from the equation …

A

specific wavelength;

epsilon = A/cl

25
Q

Beer lambert equation:

…

A

A = epsilon * b * c

26
Q 
Beer lambert: 
A = epsilon * b * c 
A = ... 
epsilon = ... 
b = ... 
c = ...
A

absorbance;
molar absorption coefficient;
path length;
concentration

27
Q

RT lab:

260/280 ratio = …; DNA is …, RNA is …

A

“purity” of nucleic acids;

  1. 8;
  2. 0
28
Q

RT lab:

260/230 ratio = second measure of …; range … - …

A

“purity”;

1.8 - 2.2

29
Q

PCR lab:

- PCR = …

A

polymerase chain reaction

30
Q

PCR lab:
PCR - rapid … of specific sequences of DNA
- bacteria vs. PCR - … vs …

A

amplification;

days; hours

31
Q 
PCR lab: 
1983 - kary mullis 
- sickle cell anemia mutation; sanger sequencing --> ... + ... 
- later added ...
- 1993 Nobel in chem
A

DNA polymerase; 2 oligo primers;

thermal cycling

32
Q 
PCR lab: 
PCR components: 
1. ... 
2. ... 
3. ... 
4. ... 
5...
A 
template; 
two primers; 
polymerase; 
nucleotides; 
thermocycler
33
Q

PCR lab:

1. template: …, sequence in the template that is to be amplified = …

A

DNA;

target

34
Q

PCR lab:

2. two primers: … whose sequence is … to …

A

oligonucleotides;
complementary;
ends of target sequence

35
Q

PCR lab:

  1. polymerase: …
  2. thermocycler: originally done by transferring between …
A

DNA-dependent DNA polymerase;

water baths

36
Q

PCR lab:
cycling:
… steps per cycle, …-… cycles

A

three;

25-35

37
Q 
PCR lab: 
cycling steps: 
1. ... 
2. ... 
3. ...
A

denature;
anneal;
elongate

Biotech (17 decks)

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