Genome Sequencing - Human Genome Project

Question 1

when did the human genome project begin?

Answer 1

October 1, 1990

Question 2

how long did this project take?

Answer 2

13 years

Question 3

who funded it?

Answer 3

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

Question 4

what DNA was used?

Answer 4

human DNA that came from several volunteers

Question 5

what were the goals of the human genome project?

Answer 5

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

Question 6

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

Answer 6

• February 2001

- 2003

Question 7

when was the final draft published?

Answer 7

2006

Question 8

how big is the human genome?

Answer 8

3 billion bp in length

Question 9

high-throughput sequencing

Answer 9

ability to rapidly sequence large amounts of DNA

Question 10

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

Answer 10

1) fluorescently labeled nucleotides and detectors

2) parallel sequencing

Question 11

next generation sequencing technologies

Answer 11

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

Question 12

pyrosequencing

Answer 12

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

Question 13

pyrosequencing steps

Answer 13

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

Question 14

in pyrosequencing, what happens after droplets are formed?

Answer 14

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

Question 15

what does the picoliter plate do?

Answer 15

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

Question 16

what is pyrosequencing an example of?

Answer 16

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

Question 17

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

Answer 17

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

Question 18

comparative genomics

Answer 18

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