Human Genome Project

Question 1

Human Genome Project

Answer 1

Was a commitee that sought to sequence the human genome, and eventually of other organism

Question 2

Who did the HGP?

Answer 2

1988 by the US national academy

Question 3

Original issues with the HGP?

Answer 3

Ethical issues, low computing power and vast amoutn of genes where sanger sequencing was limited.

Question 4

What were the two groups working on the HGP?

Answer 4

International HGP(HGS approach)
Celera Genomes(WGS approach)

Question 5

Hierarchical Genome Sequencin

Answer 5

Analysis entireity of the human genome in a cell at a single time.

Question 6

What did HGS use?

Answer 6

RE to cleave human genome ito 200,000 fragments and BAC

Question 7

Process of HGS

Answer 7

Break genome into smaller fragments(100,000NT pairs)
Ordered clone maps(comparison of restriction enzyme cleavage sites in a given BAC clone
Using GE to sequence ordered clonse, generating whole genom using PCR
COmputs alligning sequence to figure out where the chromsomes go

Question 8

What does WGS sequencing use instead?

Answer 8

Directly taking DNA into short fragments, ligated into plasmids

Question 9

Process of WGS

Answer 9

Ligating into fragments, each fragment sequenced, ordered in a computer softwar

Question 10

Why was WGS slower?

Answer 10

Computing power required due to not physically ordering the framgnets

Question 11

Differences of WGS and HGS

Answer 11

Hgs physically ordered whislt WGS did not

Question 12

HGP by 2003?

Answer 12

90% of human genom sequence

Question 13

Why was 100% sequencing hard?

Answer 13

Lots of repetetive sequences hard to allign together(especially with sanger sequencing)

Question 14

When was HGP completed?

Answer 14

2022 due to 2nd and 3rd generation sequencing

Question 15

Why were newer techniques better in sequencing?

Answer 15

Not requiring DNA libraries and use of PCR instead of BAC

Question 16

Illumnia Sequencing

Answer 16

This prepares DNA libraries by binding DNA to solid surface then amplification by PCR

Question 17

What is sequencing done with?

Answer 17

dNTP and fluorophores, so each Polymerase NT addition the machine reads light emitted, detemrining order of each dNTP

Question 18

Light emission depending on NT joined…

Answer 18

If NT joined to growing strand, PPI is released causing a flash of light, whilst if not complemnetary, no reaciton thus no flash.

Question 19

Ion Torrent Sequncing

Answer 19

Uses welds containing beads coated each with DNA, each with a different sequence

Question 20

Differnce between Ion Torrent Sequencing and Illumnia Sequencing?

Answer 20

Everytime dNTP is added, machine reads change in PH instead of fluorophores, as each dNTP added releases a proton

Question 21

Types of 2nd generation sequencing…

Answer 21

Illumnia and Ion Torrent Sequencing

Question 22

Examples of 3rd generation sequencing?

Answer 22

PACbio and Oxford Nanopores

Question 23

What can a genome sequence be used for?

Answer 23

Analysis of gene mutaiton for cancer tpyes
Analysis of parasitic pathways using known sequences of genes in humans
Identify genes in human genome compare with other species for eovlutonary genetics

Question 24

How are protein coding genes ID?

Answer 24

A protein has a reading frame with start/stop codon
Promoter sequences
RNA-splicing sites(introns/exons)

Question 25

The point of the 1000 genome projcct in 2008?

Answer 25

Between two humans, 99% of genome is identical, so studying variation to identify susceptibility to diseases

Question 26

Where HAS genome sequencing been used?

Answer 26

Cancer genomics, identification of mutations, in agriculture for crop seqeuncing, forensics, microbiomes and microbes