Module 2 Part 1 Flashcards

1
Q

define euchtomatic sequence

A

genomes that are easy to transcribe

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2
Q

what is the human genome project

A

international endeavour that resulted in the sequencing of the human genome to 99.99% accuracy

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3
Q

how long did the human genome project take

A

1990- april 2003

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4
Q

cost of genome project then vs now

A

$100 mil to $1000 USD

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5
Q

what type of sequencing did the human genome start with

A

sanger sequencing!

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6
Q

what type of technology performs DNA sequencing

A

Polymerase Chain Reaction (PCR)

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7
Q

why is DNA called DEOXYribonucleic acid?

A

DNA lacks the -OH group on C2

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8
Q

where is the phosphodiester bond located

A

between C5 of the 5’ nucleotide and C3 of the 3’ nucleotide- mediated by DNA polymerase

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9
Q

what do dideoxynucleotides lack?

A

ddNTPs lack the -OH group on C2 and C3

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10
Q

what does removing the OH group do?

A

it means no phosphodiester bond which means elongation stops

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11
Q

other names for sanger sequencing

A

-chain termination
-dideoxy sequencing method

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12
Q

key steps of sanger sequencing

A
  • primer made to known sequence
  • shear template into smaller fragments
  • sequence reaction using low concentration of ddNTPs
  • newly synthesized DNA terminated by the random incorporation of ddNTPs
  • DNA separated by size using gel electropheresis
    (small fragments first)
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13
Q

how many genes at once in sanger sequencing?

A

one

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14
Q

describe whole genome sequencing process

A

notes**

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15
Q

describe targeted panel sequencing

A
  • 40 to 400 genes
  • high coverage
  • search bias
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16
Q

describe whole exome sequencing

A

-22 000 genes
- intermediate coverage
- only sequencing exomes for certain gene

17
Q

describe whole genome sequencing

A
  • lower coverage
  • all genes, translocations and non coding DNA
  • pro: cover whole genome
  • con: no deep coverage per gene for higher accuracy
18
Q

define coverage

A

how many times a DNA region is sequenced

19
Q

how do DNA microarrays work

A

-DNA to mRNA to protein through gene expression
-mRNA to cDNA through reverse transcription

20
Q

what type of genes are transcribed into mRNA

A

protein coding genes

21
Q

what does it mean if cDNA hybridizes

A

the gene is expressed

22
Q

what is whole exome sequencing (WES)

A
  1. protein coding regions are captured and isolated using DNA microarray techniques
  2. non coding regions are washed away prior to sequencing
  3. sequencing is only performed on coding DNA
23
Q

what is first generation sequencing

A
  • sanger sequencing
  • one gene/time
  • DNA fragments: 500-1000 bp; amplification required
  • time to sequence whole genome is 10 years
  • expensive
  • highly accurate
24
Q

define 2nd generation sequencing

A
  • next generation sequencing
  • massively parallel sequencing
  • DNA fragments: 100-500 bp; amplification required
  • time to sequence genome= 1 week
  • less accurate than sanger sequencing due to coverage
25
Q

define 3rd generation sequencing

A
  • next next generation sequencing
  • massively parallel sequencing
  • DNA fragments> 1000 bp; amplification not required
  • time to sequence= 1 hour
26
Q

define ancestory testing

A

uses genotyping that identifies single nucleotide polymorphisms unique to certain populations

27
Q

define SNP

A

single nucleotide polymorphism
- common variation at single position in DNA sequence
- may lead to variations in amino acid sequence and subsequent protein

28
Q

what are the different types of genetic testing

A
  • karyotying
  • fluorescence in situ (FISH)
  • sequencing
29
Q

define ancestry

A

a person’s line of descent; their background

30
Q

define ethnicity

A

includes a persons geographical and cultural ties to their background

31
Q
A