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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

why drinking excess alcohol causes thirst and dehydration
uwikuzo Reply
what is reproduction
smart Reply
it is d act of bringing young ones to life
Oyebanji
to ensure survival of a species🚴‍♀️
Michelle
what is a genotype
Collins
what is hazardous
smart
a cell is the smallest unit of a living thing. so we all have cell
smart
It is the formation of a zygote resulting from the fusion of the sperm cell with the ovum.Thus,this results in the production of new species which are genetically dissimilar from their parent cells.
Pallavi
what is size of cell
Mohd Reply
what is size of Hart
Mohd
nanometers=um sign thingie
Michelle
microns=nanometers
Michelle
monomers and polymers of nucleic acids?
Jyrl Reply
dna and rna involvement
Michelle
give me the elements of the soil
Iguma Reply
Air, water, organic matter, inorganic matter
Tshwano
soil water humus air
Veronicah
yap
Beloved
silica, iron
Patience
potassium, sulfur, calcium, carbon
Michelle
what is cell
iyaji Reply
A cell is a smallest fundamental unit of a living organisms.
Kem
the basic structural and functional unit of life
Patience
what is size of cell
Mohd
all things are made up of.....all things cannot exist without pre-exisiting cells...check out the 16th ce tury to learn more about microscope use and cells. i will give a hint: Mr. L.
Michelle
we are all made of cells
Michelle
Nutrition - sensitive intervention
Therowda Reply
what does the sori In fern mean
arhin Reply
biology is the study .exactly what is life?
Jerry
i'm sorry , the study of life
Jerry
how can U identify a person through his blood
Frankyx Reply
through genetic fingerprinting where specific DNA sequences in a person genome can be identified.
Pallavi
by help of genetics and DNA test
Meerbadini
can it also be detected using an RNA test
Frankyx
environmental biology
Ojesola Reply
what is phytoplankton
Ojesola
What is anabolism
Treasure Reply
the break down of substance
Aunty
how many teeth has an adult person
TSEGAY Reply
36
Treasure
32
Favour
32
Kamaluddeen
32 teeth
Meerbadini
32
OLAMIDE
32 (or 36 including the wisdom teeth)
Hailey
32
Sani
26?🤓
Michelle
what is homoestastis
Ayo Reply
hypothalamus negative feedback vs. postive feedback systems.
Michelle
it is the maintenance of a steady internal environment.it is controlled largely by the brain especially the hypothalamus.
Oyebanji
What is a cell
Eric Reply
A cell is the building block of all organisms
Millicent
yes it is , it also helps in functions .
Joezman
cell is structural & functional unit of oraganisms
Meerbadini
it is a building blocks of a living organism
Joezman
it is the basic unit of life in living organisms
Nabwonso
it is the basic unite of life in living organisms
Nabwonso
it is the fundamental unit of life
Adewoye
it is the structural and functional unit of life
Ojesola
Why cells are classify as living and non living things
Millicent Reply
due to such as virus which acts as a living cell inside the cell of an organisms and out side of cell in form of crystals
Meerbadini
because there are living cells which performs complex processes such as reproduction and metabolism then there are dead cells like the top most part of the epidermis or the bark
irish
because there are living cells which performs complex function such as reproduction and metabolism
David
cause otherwise you could call fire alive
Marissa

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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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