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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

Simple term of science
Palesa Reply
what does it mean
gopal
it's means what do u know about biology?
Phathu
what is immunisation
Melysa
what is the biology? what do you know about biology
Phathu Reply
biology is the study of living organisms, divided into many specialized fields that cover their morphology, physiology, anatomy, behavior, origin, and distribution.
Julia
The study of all aspects of life. The study of all living organisms (such as animal cells and plant cells) in greater detail (their structure and how they function). It's a very broad science.
juanita
what is prokaryotic
Bhaskar Reply
what is pathogens
Bhaskar
transistion metals....
Wasik Reply
Why study ecology
Amos Reply
What name is given to group 8metals on a periodic table
Amos
what is evolution
Elia Reply
Is the gradual change of something it can be either organisms
Amos
which of the following statements about the moss life cycle is false?
Israel Reply
posterior lobe of pitutary contains what?
MR Reply
What if vincristine and colchicibe disrupt mitosis by binding to tubulin
Rohith Reply
A plant in the understory of a forest displays a segmented stem and slender leaves arranged in a whorl. It is probably....
Israel Reply
How did the development of a vascular system contribute to the increase in size of plants?
Israel
If a flower lacked a megasporangium and microsporangium, what type of gametes would not form?
Israel
Seed plants are.. A. all homosporous B. mostly homosporous with some heterosporous C. mostly heterosporous with some homosporous D. all heterosporous
Israel
Besides the seed, what other major structure diminishes a plant's reliance on water for reproduction?
Israel
what role did the adaptations of seed and pollen play in the development and expansion of seed plants?
Israel
Some cycads are considered endangered species and their trade is severely restricted. Customs officials stop suspected smugglers who claim that the plants in their possession are palm trees, not cycads. How would a botanist distinguish between the two types of plants?
Israel
What are the two structures that allow angiosperms to be the dominant form of plant life in most terrestrial ecosystems?
Israel
how are carbohydrates,proteins and fats formed from triose phosphate
fonyuy Reply
Why does the actin filament only move in one direction? Describe in great detail.
Lashonda Reply
Two events happen when calcium binds to troponin.
Lashonda
which of the following statements about plant division is false?
Israel Reply
😂😂😂 i dont know how i reached here
esther
What are the events that occur in each phase of interphase
Hazey Reply
what is photosynthesis
Victor Reply
The process plants use to convert sunlight into food (energy).
juanita
Some other organisms use
juanita
types of photosynthesis
Victor
ps1 and ps2
smritilekha

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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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