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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 parasitic movement
Emmanuel Reply
Parasitic movement is a problem for all of us. So is its companion, parasitic tension. Parasitic movement is the excess contraction of muscles that you don't actually need to complete an action.
freya
what are eukaryotic cells
Thiza Reply
what is the mean of pair of chromosomes
Kazula Reply
hi
Lagos
23 haploid and 23diploid
Patson
how are you studying in this quarantine? .. how are you keeping yourselves motivated?
sivajijadhav @815.com
good morning guyz
Joelia
morning
Kazula
hi
Justin
Good
Angela
tell me if you know what can be used...than reading pls hint me pls 🙏🙏🙏
Angela
what is the important of sex
Aremu Reply
why did human being need sex?
Aremu
because he/she have feelings
Chripine
reproduction...to make more
Yazi
due to active harmon
Manish
One important of sex is to reproduce
Emma
to ensure the countinuty of life
Yusuf
all of you are right
Edith
what is momentum
Asiya Reply
The strength or force that allows something to continue or grow stronger or faster as time pass
Emma
What is Centripetal Force?
Justin
centrepital force is the inward force required to keep a body moving with constant speed in a circular path
Yusuf
what is the test for protein
Takii Reply
List four condition necessary for seed germination
Tedeka Reply
water,air(oxygen),light,temperature
Hassan
water, light, oxygen and temperature
lilchris
water, oxygen, light temperature
Mike
water oxygen light and temperature
John
importance of biology
Alabina Reply
importance of boilogy
Alabina
what is soil
Amina Reply
soil is the upper part of the earth
Alabina
what is importance of studying biology
Alabina
soil is the uppermost layer of the earth on which plant grows
Yusuf
soil is defined as the thin surface of the upper most layer of the earth crust on which plants grow
Thanni
soil is the upper part of the earth which plants grow on
Emma
differences between euglenoid and amoeboid
Grace Reply
what are the difference between aerobic and anaerobic respiration?
Maxwell Reply
Aerobic respiration involves the use of oxygen whiles anaerobic respiration does not involve the use of oxygen
Quabena
what is assmilation
Lucy Reply
what is cell
Manish Reply
cell is the structural and functional unit of life or living things
hamid
where anaerobic respiration occurre?
Manish
in cell
Manish
in cells?
Manish
where anaerobic respiration occurre in cell?
Manish
what's phloem?
Gift Reply

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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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