Difference between pages "Essay !4 - About Sequencing Code : KSI0004" and "Essay !5 - Genome Project Code: KSI0005"

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<p>S1.4 Genomics Essay #4</p>
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<p>Essay 5 &ndash; Human Genome Project</p>
  
<p><a href="http://biolecture.org/index.php/Essay_!4_-_About_Sequencing_Code_:_KSI0004"><u>http://biolecture.org/index.php/Essay_!4_-_About_Sequencing_Code_:_KSI0004</u></a></p>
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<p>and Ulsan Genome Project by Prof. Jong Bhak.</p>
 
 
<p>&nbsp;</p>
 
 
 
<p>Essay 4 &ndash; About Sequencing</p>
 
  
 
<p>Sangin Kim</p>
 
<p>Sangin Kim</p>
Line 11: Line 7:
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>
  
<p>In genetics and biochemistry, sequencing means to determine the primary structure of an unbranched biopolymer. Sequencing results in a symbolic linear depiction known as a sequence which succinctly summarizes much of the atomic-level structure of the sequenced molecule.</p>
+
<p>The Human Genome Project (HGP) was an international scientific research project with the goal of determining the sequence of nucleotide base pairs that make up human DNA, and of identifying and mapping all of the genes of the human genome from both a physical and a functional standpoint.</p>
 
 
<p>&nbsp;</p>
 
 
 
<p>DNA sequencing is the process of determining the nucleotide order of a given DNA fragment. This technique uses sequence-specific termination of a DNA synthesis reaction using modified nucleotide substrates. However, new sequencing technologies such as pyrosequencing are gaining an increasing share of the sequencing market. More genome data are now being produced by pyrosequencing than Sanger DNA sequencing. Pyrosequencing has enabled rapid genome sequencing. Bacterial genomes can be sequenced in a single run with several times coverage with this technique.</p>
 
 
 
<p>The sequence of DNA encodes the necessary information for living things to survive and reproduce. Determining the sequence is therefore useful in fundamental research into why and how organisms live, as well as in applied subjects. Because of the key importance DNA has to living things, knowledge of DNA sequences are useful in practically any area of biological research. For example, in medicine it can be used to identify, diagnose, and potentially develop treatments for genetic diseases. Similarly, research into pathogens may lead to treatments for contagious diseases. Biotechnology is a burgeoning discipline, with the potential for many useful products and services.</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>1.1 sanger sequencing</p>
 
  
<p>In chain terminator sequencing (Sanger sequencing), extension is initiated at a specific site on the template DNA by using a short oligonucleotide &#39;primer&#39; complementary to the template at that region. The oligonucleotide primer is extended using a DNA polymeraase an enzyme that replicates DNA. Included with the primer and DNA polymerase are the four deoxynucleotide bases (DNA building blocks), along with a low concentration of a chain terminating nucleotide (most commonly a di-deoxynucleotide). Limited incorporation of the chain terminating nucleotide by the DNA polymerase results in a series of related DNA fragments that are terminated only at positions where that particular nucleotide is used. The fragments are then size-separated by electrophoresis in a slab polyacrylamide gel, or more commonly now, in a narrow glass tube (capillary) filled with a viscous polymer.</p>
+
<p>The process of identifying the boundaries between genes and other features in a raw DNA sequence is called genome annotation and is in the domain of bioinformatics. The genome published by the HGP does not represent the sequence of every individual&#39;s genome. It is the combined mosaic of a small number of anonymous donors, all of European origin. The HGP genome is a scaffold for future work in identifying differences among individuals. Subsequent projects sequenced the genomes of multiple distinct ethnic groups, though as of today there is still only one &quot;reference genome.&quot; The human genome has significantly more segmental duplications (nearly identical, repeated sections of DNA) than had been previously suspected. At the time when the draft sequence was published fewer than 7% of protein families appeared to be vertebrate specific.</p>
  
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>
  
<p>&nbsp;</p>
+
<p>These techniques include:</p>
  
<p>1.2 Pyrosequencing</p>
+
<p>DNA Sequencing</p>
  
<p>Pyrosequencing which was developed by P&aring;l Nyr&eacute;n and Mostafa Ronaghi, has been commercialized by Biotage (for low-throughput sequencing) and 454 Life Sciences (for high-throughput sequencing). The latter platform sequences roughly 100 megabases [now up to 400 megabases] in a seven-hour run with a single machine. In the array-based method (commercialized by 454 Life Sciences), single-stranded DNA is annealed to beads and amplified via EmPCR. These DNA-bound beads are then placed into wells on a fiber-optic chip along with enzymes which produce light in the presence of ATP. When free nucleotides are washed over this chip, light is produced as ATP is generated when nucleotides join with their complementary base pairs. Addition of one (or more) nucleotide(s) results in a reaction that generates a light signal that is recorded by the CCD camera in the instrument. The signal strength is proportional to the number of nucleotides, for example, homopolymer stretches, incorporated in a single nucleotide flow.</p>
+
<p>The Employment of Restriction Fragment-Length Polymorphisms (RFLP)</p>
  
<p>&nbsp;</p>
+
<p>Yeast Artificial Chromosomes (YAC)</p>
  
<p>1.3 Large scale sequencing</p>
+
<p>Bacterial Artificial Chromosomes (BAC)</p>
  
<p>Whereas the methods above describe various sequencing methods, separate related terms are used when a large portion of a genome is sequenced. Several platforms were developed to perform exome sequencing (a subset of all DNA across all chromosomes that encode genes) or whole genome sequencing (sequencing of the all nuclear DNA of a human).</p>
+
<p>The Polymerase Chain Reaction (PCR)</p>
  
<p>&nbsp;</p>
+
<p>Electrophoresis</p>
  
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>
  
<p>Taxonomy</p>
+
<p>&lt; The Ulsan Genome project &gt; - News article.</p>
 
 
<p>&bull;Sequencing: determining the precise order of nucleotides in a DNA or RNA molecule</p>
 
 
 
<p>&bull;Sanger dideoxy method</p>
 
  
<p>&bull;Invented by Nobel Prize winner Fred Sanger</p>
+
<p>Genome Korea in Ulsan Launched</p>
  
<p>&bull;Dideoxy analogs of dNTPs used in conjunction with dNTPs</p>
+
<p>- The largest scale Personal Genome Project in Korea to sequence 10,000 people and patients -</p>
  
<p>&bull;Analog prevents further extension of DNA chain</p>
+
<p>ULSAN, KOREA - Nov 25, 2015 -</p>
  
<p>&bull;Bases are labeled with radioactivity</p>
+
<p>The Ulsan 10,000 Genome Project, entitled the “Genome Korea in Ulsan” has been launched in Ulsan Metropolitan City on the 25th of Nov. 2015. The consortium includes the Ulsan Metropolitan City, Ulsan National Institute of Science and Technology (UNIST), Ulsan University Hospital, and the University of Ulsan.</p>
  
<p>&bull;Gel electrophoresis is then performed on products</p>
+
<p>This is a large-scale publicly-funded genome project in Korea, with the estimated funding of ~25 million USD by 2019(not fully acquired yet). The goal is to map complete genomic diversity of Koreans, constructing standardized gene variation database, detecting rare genetic mutations, and providing well-annotated full genome information for growing genomic industry of Korea. The consortium will seek necessary funding from public and private sectors to achieve its goal of sequencing all the Koreans in the next decades. The initial 10,000 samples will be collected from both healthy people and immunocompromised people.</p>
  
<p>&bull;Large-scale sequencing projects have led to automated DNA sequencing systems</p>
+
<p>The project’s practical aim is to develop an industrial foundation in genomics for future biomedical industry. Ulsan, known as the capital of Korean industrialization, has a well established industrial infrastructure. The consortium will facilitate developing new sequencing and analysis technologies to achieve personalized medicine in Korea. This project is complementary to Korean government’s on-going Multi-ministry Genomics Initiative which has started in 2013 with a total sum of 500 million USD for 8 years to carry out human, agricultural, and medical genomics projects.</p>
  
<p>&bull;Based on Sanger method</p>
+
<p>Genome Korea is in collaboration with Harvard Medical School’s Personal Genome Project (PGP), led by Professor George Church who developed key genome sequencing and editing technologies for decades. UNIST and Harvard Medical School will sign an MOU for the Ulsan 10,000 genome project.</p>
  
<p>&bull;Radioactivity replaced by fluorescent dye</p>
+
<p>Genome Korea is a participatory project where volunteers donate blood samples and personal and clinical information. Korean PGP project, led by Prof. Jong Bhak at UNIST have already published over 50 high quality individual genomes with the Korean reference genome assembly, funded by Korean government. Ulsan’s 10,000 genome is the first large scale public project that will expand to the whole Korean population which is similar to 100,000 UK genome and US president Obama’s 1 million genome project.</p>
  
<p>&bull;Virtually all genomic sequencing projects use shotgun sequencing</p>
+
<p>Ulsan mayor, Mr. Ki-hyun Kim, emphasized the significance of Genome Korea in Ulsan project by addressing “We aim to make Ulsan as the hub of genomic industry in Asia and beyond by linking it to diagnostic and therapeutic medical industry as a key Korean economic industrialization driving force”.</p>
  
<p>&bull;Entire genome is cloned, and resultant clones are sequenced</p>
+
<p>UNIST president, Prof. Mooyoung Jung has an ambitious plan to make this Ulsan 10,000 genome project, by raising the technology level to “the world top level innovative research by analyzing 10,000 people genomes at UNIST”.</p>
  
<p>&bull;Much of the sequencing is redundant</p>
+
<p>“Korea&#39;s aging population is growing at a rapid pace. We, therefore, need genome industry to lower the medical cost and prevent national scale infectious disease endemic analyzing genomes and associated omics information. This must be accompanied by the commercialization of the technologies” and “Genome project can function as the seed of future biomedical revolution in business and society”, says Prof. Jong Bhak, the lead researcher of this project.</p>
  
<p>&bull;Generally 7- to 10-fold coverage</p>
+
<p>Ulsan is the most industrialized city in Korea which hosts global business cooperations such as Hyundai, SK, and Samsung. Ulsan’s main industry has been mostly heavy industrials such as car manufacturing, ship building, and oil refining. Currently, Ulsan plans to develop new high-tech industries such as biomedical devices, reagents, new materials, energy storage, and information technologies. Ulsan 10K genome is a part of such an effort to recruit skilled labor and highly value-added business entities.</p>
  
<p>&bull;Computer algorithms are used to look for replicate sequences and assemble them</p>
+
<p>UNIST is a new science and technology university in Korea established by the government in 2009.</p>
  
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>
  
<p>NGS(Next generation sequencing)</p>
+
<p>References</p>
 
 
<p>&bull;Second-generation DNA sequencing</p>
 
  
<p>&bull;Generates data 100x faster than Sanger method</p>
+
<p>1.https://en.wikipedia.org/wiki/Human_Genome_Project</p>
  
<p>&bull;Massively parallel methods</p>
+
<p>2. http://bme.unist.ac.kr/launch-of-the-genome-korea-in-ulsan/?lang=ko</p>
 
 
<p>&bull;Large number of samples sequenced side by side</p>
 
 
 
<p>&bull;Uses increased computer power and miniaturization</p>
 
 
 
<p>&bull;454 Life Sciences pyrosequencing</p>
 
 
 
<p>&bull;Illumina/Solexa sequencing</p>
 
 
 
<p>&bull;SOLID/Applied Biosystems method</p>
 
 
 
<p>&bull;454 sequencing system</p>
 
 
 
<p>&bull;DNA is broken into small segments</p>
 
 
 
<p>&bull;DNA is amplified using polymerase chain reaction (PCR)</p>
 
 
 
<p>&bull;Light is released each time a base is added to DNA strand</p>
 
 
 
<p>&bull;Instrument actually measures release of light</p>
 
 
 
<p>&bull;Can handle only short stretches of DNA strand</p>
 
 
 
<p>&bull;Third-generation DNA sequencing</p>
 
 
 
<p>&bull;Sequencing of single molecules of DNA</p>
 
 
 
<p>&bull;HeliScope Single Molecule Sequencer</p>
 
 
 
<p>&bull;Single-stranded DNA fragments attached in array on glass slide</p>
 
 
 
<p>&bull;Complementary strand synthesized</p>
 
 
 
<p>&bull;Fluorescent tags monitored on microscope</p>
 
 
 
<p>&bull;Computer assembles fragments into sequence</p>
 
 
 
<p>&bull;Third-generation DNA sequencing</p>
 
 
 
<p>&bull;Pacific Biosciences SMRT</p>
 
 
 
<p>&bull;Single Molecule Real Time sequencing</p>
 
 
 
<p>&bull;Reactions carried out in nanocontainers (zero mode wave guides)</p>
 
 
 
<p>&bull;Single-stranded DNA fragments attached</p>
 
 
 
<p>&bull;Complementary strand synthesized</p>
 
 
 
<p>&bull;Fluorescent tags monitored</p>
 
 
 
<p>&bull;Computer assembles fragments into sequence</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>&bull;Fourth-generation DNA sequencing</p>
 
 
 
<p>&bull;Optical detection no longer used</p>
 
 
 
<p>&bull;Ion torrent semiconductor sequencing</p>
 
 
 
<p>&bull;Measures release of protons whenever a deoxyribonucleotide is added</p>
 
 
 
<p>&bull;Silicon chip measures &quot;pH&quot;</p>
 
 
 
<p>&bull;Extremely fast</p>
 
 
 
<p>&bull;Less expensive</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>&bull;Fourth-generation DNA sequencing</p>
 
 
 
<p>&bull;Oxford Nanopore Technologies system</p>
 
 
 
<p>&bull;Passes DNA through nanoscale biological pores</p>
 
 
 
<p>&bull;Detector measures change in electric current</p>
 
 
 
<p>&bull;Extremely fast</p>
 
 
 
<p>&bull;Measures long chains of DNA</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>2. RNA sequencing</p>
 
 
 
<p>RNA is less stable in the cell, and also more prone to nuclease attack experimentally. As RNA is generated by transcription from DNA, the information is already present in the cell&#39;s DNA. However, it is sometimes desirable to sequence RNA molecules. While sequencing DNA gives a genetic profile of an organism, sequencing RNA reflects only the sequences that are actively expressed in the cells. To sequence RNA, the usual method is first to reverse transcribe the RNA extracted from the sample to generate cDNA fragments. This can then be sequenced as described above. The bulk of RNA expressed in cells are ribosomal RNAs or small RNAs, detrimental for cellular translation, but often not the focus of a study. This fraction can fortunately be removed in vitro, however, to enrich for the messenger RNA, also included, that usually is of interest. Derived from the exons these mRNAs are to be later translated to proteins that support particular cellular functions. The expression profile therefore indicates cellular activity, particularly desired in the studies of diseases, cellular behaviour, responses to reagents or stimuli. Eukaryotic RNA molecules are not necessarily co-linear with their DNA template, as introns are excised. This gives a certain complexity to map the read sequences back to the genome and thereby identify their origin. For more information on the capabilities of next-generation sequencing applied to whole transcriptomes see: RNA-Seq and MicroRNA Sequencing.</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>3. Protein sequcing</p>
 
 
 
<p>If the gene encoding the protein is known, it is currently much easier to sequence the DNA and infer the protein sequence. Determining part of a protein&#39;s amino-acidsequence (often one end) by one of the above methods may be sufficient to identify a clone carrying this gene.</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>4. Polysacharride sequencing</p>
 
 
 
<p>Though polysaccharides are also biopolymers, it is not so common to talk of &#39;sequencing&#39; a polysaccharide, for several reasons. Although many polysaccharides are linear, many have branches. Many different units (individual monosaccharides) can be used, and bonded in different ways. However, the main theoretical reason is that whereas the other polymers listed here are primarily generated in a &#39;template-dependent&#39; manner by one processive enzyme, each individual join in a polysaccharide may be formed by a different enzyme. In many cases the assembly is not uniquely specified; depending on which enzyme acts, one of several different units may be incorporated. This can lead to a family of similar molecules being formed. This is particularly true for plant polysaccharides. Methods for the structure determination of oligosaccharides and polysaccharidesinclude NMR spectroscopy and methylation analysis.</p>
 
 
 
<p>&nbsp;</p>
 
 
 
<p>&nbsp;</p>
 
  
<p>Reference</p>
+
<p>3.http://www.yonhapnews.co.kr/bulletin/2015/11/25/0200000000AKR20151125116000057.HTML</p>
  
<p>1. https://www.youtube.com/watch?v=jFCD8Q6qSTM</p>
+
<p>4. https://www.genome.gov/12011238/an-overview-of-the-human-genome-project/</p>
  
<p>2. Brook biology of microorganisms 14th edition.</p>
+
<p>5. http://www.bio-itworld.com/Press-Release/Genome-Korea-in-Ulsan-Launched/</p>
  
 
<p>&nbsp;</p>
 
<p>&nbsp;</p>

Latest revision as of 03:34, 3 December 2016

Essay 5 – Human Genome Project

and Ulsan Genome Project by Prof. Jong Bhak.

Sangin Kim

 

The Human Genome Project (HGP) was an international scientific research project with the goal of determining the sequence of nucleotide base pairs that make up human DNA, and of identifying and mapping all of the genes of the human genome from both a physical and a functional standpoint.

The process of identifying the boundaries between genes and other features in a raw DNA sequence is called genome annotation and is in the domain of bioinformatics. The genome published by the HGP does not represent the sequence of every individual's genome. It is the combined mosaic of a small number of anonymous donors, all of European origin. The HGP genome is a scaffold for future work in identifying differences among individuals. Subsequent projects sequenced the genomes of multiple distinct ethnic groups, though as of today there is still only one "reference genome." The human genome has significantly more segmental duplications (nearly identical, repeated sections of DNA) than had been previously suspected. At the time when the draft sequence was published fewer than 7% of protein families appeared to be vertebrate specific.

 

These techniques include:

DNA Sequencing

The Employment of Restriction Fragment-Length Polymorphisms (RFLP)

Yeast Artificial Chromosomes (YAC)

Bacterial Artificial Chromosomes (BAC)

The Polymerase Chain Reaction (PCR)

Electrophoresis

 

< The Ulsan Genome project > - News article.

Genome Korea in Ulsan Launched

- The largest scale Personal Genome Project in Korea to sequence 10,000 people and patients -

ULSAN, KOREA - Nov 25, 2015 -

The Ulsan 10,000 Genome Project, entitled the “Genome Korea in Ulsan” has been launched in Ulsan Metropolitan City on the 25th of Nov. 2015. The consortium includes the Ulsan Metropolitan City, Ulsan National Institute of Science and Technology (UNIST), Ulsan University Hospital, and the University of Ulsan.

This is a large-scale publicly-funded genome project in Korea, with the estimated funding of ~25 million USD by 2019(not fully acquired yet). The goal is to map complete genomic diversity of Koreans, constructing standardized gene variation database, detecting rare genetic mutations, and providing well-annotated full genome information for growing genomic industry of Korea. The consortium will seek necessary funding from public and private sectors to achieve its goal of sequencing all the Koreans in the next decades. The initial 10,000 samples will be collected from both healthy people and immunocompromised people.

The project’s practical aim is to develop an industrial foundation in genomics for future biomedical industry. Ulsan, known as the capital of Korean industrialization, has a well established industrial infrastructure. The consortium will facilitate developing new sequencing and analysis technologies to achieve personalized medicine in Korea. This project is complementary to Korean government’s on-going Multi-ministry Genomics Initiative which has started in 2013 with a total sum of 500 million USD for 8 years to carry out human, agricultural, and medical genomics projects.

Genome Korea is in collaboration with Harvard Medical School’s Personal Genome Project (PGP), led by Professor George Church who developed key genome sequencing and editing technologies for decades. UNIST and Harvard Medical School will sign an MOU for the Ulsan 10,000 genome project.

Genome Korea is a participatory project where volunteers donate blood samples and personal and clinical information. Korean PGP project, led by Prof. Jong Bhak at UNIST have already published over 50 high quality individual genomes with the Korean reference genome assembly, funded by Korean government. Ulsan’s 10,000 genome is the first large scale public project that will expand to the whole Korean population which is similar to 100,000 UK genome and US president Obama’s 1 million genome project.

Ulsan mayor, Mr. Ki-hyun Kim, emphasized the significance of Genome Korea in Ulsan project by addressing “We aim to make Ulsan as the hub of genomic industry in Asia and beyond by linking it to diagnostic and therapeutic medical industry as a key Korean economic industrialization driving force”.

UNIST president, Prof. Mooyoung Jung has an ambitious plan to make this Ulsan 10,000 genome project, by raising the technology level to “the world top level innovative research by analyzing 10,000 people genomes at UNIST”.

“Korea's aging population is growing at a rapid pace. We, therefore, need genome industry to lower the medical cost and prevent national scale infectious disease endemic analyzing genomes and associated omics information. This must be accompanied by the commercialization of the technologies” and “Genome project can function as the seed of future biomedical revolution in business and society”, says Prof. Jong Bhak, the lead researcher of this project.

Ulsan is the most industrialized city in Korea which hosts global business cooperations such as Hyundai, SK, and Samsung. Ulsan’s main industry has been mostly heavy industrials such as car manufacturing, ship building, and oil refining. Currently, Ulsan plans to develop new high-tech industries such as biomedical devices, reagents, new materials, energy storage, and information technologies. Ulsan 10K genome is a part of such an effort to recruit skilled labor and highly value-added business entities.

UNIST is a new science and technology university in Korea established by the government in 2009.

 

References

1.https://en.wikipedia.org/wiki/Human_Genome_Project

2. http://bme.unist.ac.kr/launch-of-the-genome-korea-in-ulsan/?lang=ko

3.http://www.yonhapnews.co.kr/bulletin/2015/11/25/0200000000AKR20151125116000057.HTML

4. https://www.genome.gov/12011238/an-overview-of-the-human-genome-project/

5. http://www.bio-itworld.com/Press-Release/Genome-Korea-in-Ulsan-Launched/