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By the end of this section, you will be able to:
  • Describe three types of sequencing
  • Define whole-genome sequencing

Although there have been significant advances in the medical sciences in recent years, doctors are still confounded by some diseases, and they are using whole-genome sequencing to get to the bottom of the problem. Whole-genome sequencing is a process that determines the DNA sequence of an entire genome. Whole-genome sequencing is a brute-force approach to problem solving when there is a genetic basis at the core of a disease. Several laboratories now provide services to sequence, analyze, and interpret entire genomes.

For example, whole-exome sequencing is a lower-cost alternative to whole genome sequencing. In exome sequencing, only the coding, exon-producing regions of the DNA are sequenced. In 2010, whole-exome sequencing was used to save a young boy whose intestines had multiple mysterious abscesses. The child had several colon operations with no relief. Finally, whole-exome sequencing was performed, which revealed a defect in a pathway that controls apoptosis (programmed cell death). A bone-marrow transplant was used to overcome this genetic disorder, leading to a cure for the boy. He was the first person to be successfully treated based on a diagnosis made by whole-exome sequencing. Today, human genome sequencing is more readily available and can be completed in a day or two for about $1000.

Strategies used in sequencing projects

The basic sequencing technique used in all modern day sequencing projects is the chain termination method (also known as the dideoxy method), which was developed by Fred Sanger in the 1970s. The chain termination method involves DNA replication of a single-stranded template with the use of a primer and a regular deoxynucleotide    (dNTP), which is a monomer, or a single unit, of DNA. The primer and dNTP are mixed with a small proportion of fluorescently labeled dideoxynucleotides (ddNTPs). The ddNTPs are monomers that are missing a hydroxyl group (–OH) at the site at which another nucleotide usually attaches to form a chain ( [link] ). Each ddNTP is labeled with a different color of fluorophore. Every time a ddNTP is incorporated in the growing complementary strand, it terminates the process of DNA replication, which results in multiple short strands of replicated DNA that are each terminated at a different point during replication. When the reaction mixture is processed by gel electrophoresis after being separated into single strands, the multiple newly replicated DNA strands form a ladder because of the differing sizes. Because the ddNTPs are fluorescently labeled, each band on the gel reflects the size of the DNA strand and the ddNTP that terminated the reaction. The different colors of the fluorophore-labeled ddNTPs help identify the ddNTP incorporated at that position. Reading the gel on the basis of the color of each band on the ladder produces the sequence of the template strand ( [link] ).

A deoxynucleotide consists of a deoxyribose sugar, a base, and three phosphate groups. Dideoxyribose is identical to deoxyribose except that the hydroxyl (–OH) group at the 3' position is replaced by H. A 3' hydroxyl is necessary for elongation of the DNA chain, and the chain therefore stops growing if a dideoxyribose instead of deoxyribose is incorporated into the growing chain.
A dideoxynucleotide is similar in structure to a deoxynucleotide, but is missing the 3' hydroxyl group (indicated by the box). When a dideoxynucleotide is incorporated into a DNA strand, DNA synthesis stops.

Questions & Answers

what is class bryophyta
Emefa Reply
how many stages do we have in glycolysis?
Damali
10 stages
Elisha
the presence of a membrane enclosed nuclosed is a characteristics of what
Addai Reply
eukaryotic cell
captain
hetreothalism in fungi
Lekhram Reply
there are 3 trimester in human pregnancy
ROHIN Reply
I don't know answer of this question can u help me
ROHIN
yes
Bisa
what is a cell
Fatima Reply
A cell is functional and structural unit of life.
Bisa
what is genetic
Janet Reply
I join
Janet
what are the branchas of biology
Prisca Reply
zoology, ecology
Millicent
biochemistry,cytology,herpetology...etc
R0se
genetics, microbiology,botany and embryology
Muhammad
what is a cell
Kulunbawi Reply
cell is smallest unit of life. cells are often cell the building blocks of life...
Muhammad
the first twenty element
Orapinega Reply
what are the characteristics of living things?
R0se
growth,respiration,nutrition,sensitivity, movement,irritability, excretion,death.
Obinna
What is the difference between adaptation and competition in animals
Adeyemi Reply
What is biology
Adeyemi
it is a natural science stadey about living things
Zamiil
Biology is the bronch of science which deals with the study of life is called biology
Aziz
what is the x in 300 stands for?
Ogbudu Reply
the properties of life
Clarinda Reply
response to the environment, reproduction, homeostasis, growth,energy processing etc.....
Pushpam
hello.
Daniela
hi
MacPeter
Good
Thomas
what is reproduction
Tims
Reproduction is a fundamental feature of all known life,each individual organism exist as a result of re production.....or else Multiplying...
R0se
a complete virus particle known as
Darlington Reply
These are formed from identical protein subunitscalled capsomeres.
Pushpam
fabace family plant name
Pushpam Reply
in eukaryotes ...protein channel name which transport protein ...
Pushpam Reply
in bacteria ...chromosomal dna duplicate structure called
Pushpam

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Source:  OpenStax, Biology. OpenStax CNX. Feb 29, 2016 Download for free at http://cnx.org/content/col11448/1.10
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