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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 biology
PEACE Reply
what is biology
Ysabella Reply
biology is a study of living things
PEACE
Biology is a diverse branch of science that deals with mostly living things
Emmanuel
yes
Swapnil
What happen when inhibit the transcription?
Swapnil
what is the effect of not doing sexual intercourse
SUZAN Reply
what is the mechanism of cellular respiration
Rita Reply
what is enzyme
garry Reply
They are organic catalysts that alter the rate of chemical reactions in the body.
Iyadi
what is a cell
Praize Reply
what is biology
Mordi Reply
biology is the study of living organisms and their interactions with one another and their environments
Ysabella
which of the following event does not occur during some stages of interface?
Bangha Reply
What is microfilaments
KHalid Reply
What is multicellular organisms
Ovie Reply
these are organisms with more than two cells
Bangha
the process when a male toad fertilizer a female eggs is called what?
Ahrebe Reply
Fertilization
Gyamfi
how did unicellular organisms form plants and animals or is it that different unicellular organisms formed plants and animald
YXNG Reply
yes
James
thanks
YXNG
name the components of faeces
Damali
undigested carbohydrate, fibre
Sandra
what are unicellular organisms..?
Sackson
they have only one cell
Sandra
faeces contains many undigested food materials, after the food has been digested then it will be absorbed in the blood stream for assimilation.,......... but the remains toxic materials are stored in the rectum these toxic materials are the faeces and it contains bile salts, the polysaccharides .
James
nice.
Sandra
thanks
James
unicellular organisms are the ones with only single cell.
James
thanks for your answers guys.
Sackson
Ok
Richard
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

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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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