Genome Sequencing - Human Genome Project Flashcards

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1
Q

when did the human genome project begin?

A

October 1, 1990

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2
Q

how long did this project take?

A

13 years

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3
Q

who funded it?

A

U.S. Department of Energy and the National Institutes of health

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4
Q

what DNA was used?

A

human DNA that came from several volunteers

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5
Q

what were the goals of the human genome project?

A

1) obtain a genetic linkage map of the human genome
2) obtain a physical map of the human genome
3) obtain DNA sequence of the entire human genome
4) develop technology for the management of human genome information
5) analyze the genomes of model organisms
6) develop programs focused on understanding and addressing ethical, legal and social implications of the results obtained from the human genome project
7) develop technological advances in genetic methodologies

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6
Q

when was the first draft of the nearly complete sequence published? the second?

A
  • February 2001

- 2003

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7
Q

when was the final draft published?

A

2006

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8
Q

how big is the human genome?

A

3 billion bp in length

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9
Q

high-throughput sequencing

A

ability to rapidly sequence large amounts of DNA

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10
Q

what are some advances that have made rapidly sequencing DNA possible?

A

1) fluorescently labeled nucleotides and detectors

2) parallel sequencing

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11
Q

next generation sequencing technologies

A

newer DNA sequencing tehcnologies that are more rapid and inexpensive than the dideoxy method

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12
Q

pyrosequencing

A

relies on the release of pyrophosphate, sis for the Roche 454/FLX

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13
Q

pyrosequencing steps

A

1) DNA is broken into small fragments of 300-800 bp
2) short oligonucleotides called adapters are linked to the 5’ and 3’ ends of DNA fragements
3) DNA is denatured to single strands
4) mixture of single strand DNA fragments with adaptors is called the sample library - single strands and beads attach via red adaptors (one strand per bead)
5) emulsify the beads so there is only one bead per droplet, droplets also contain pcr reagents that amplify the DNA

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14
Q

in pyrosequencing, what happens after droplets are formed?

A

each bead is placed into a well of a picoliter plate, where sequencing reagents are added to the wells

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15
Q

what does the picoliter plate do?

A

acts like a flow cell in which the beads are stuck in thew wells an pure solutions of different nucleotides can flow over them
- this is real time DNA

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16
Q

what is pyrosequencing an example of?

A

sequencing by synthesis (SBS) - involves identification of each nucleotide immediately after its incorporation into a DNA strand by DNA polymerase

17
Q

how do you know if the nucleotide has been added to a growing DNA strand?

A

pyrophosphate would have been released and the reaction would give off light

18
Q

comparative genomics

A

using information from genome projects to understand the genetic variation between different populations and evolutionary relationships among different species