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In the northern blot , another variation of the Southern blot, RNA (not DNA) is immobilized on the membrane and probed. Northern blots are typically used to detect the amount of mRNA made through gene expression within a tissue or organism sample.

Microarray analysis

Another technique that capitalizes on the hybridization between complementary nucleic acid sequences is called microarray analysis . Microarray analysis is useful for the comparison of gene-expression patterns between different cell types—for example, cells infected with a virus versus uninfected cells, or cancerous cells versus healthy cells ( [link] ).

Typically, DNA or cDNA from an experimental sample is deposited on a glass slide alongside known DNA sequences. Each slide can hold more than 30,000 different DNA fragment types. Distinct DNA fragments (encompassing an organism’s entire genomic library) or cDNA fragments (corresponding to an organism’s full complement of expressed genes) can be individually spotted on a glass slide.

Once deposited on the slide, genomic DNA or mRNA can be isolated from the two samples for comparison. If mRNA is isolated, it is reverse-transcribed to cDNA using reverse transcriptase. Then the two samples of genomic DNA or cDNA are labeled with different fluorescent dyes (typically red and green). The labeled genomic DNA samples are then combined in equal amounts, added to the microarray chip, and allowed to hybridize to complementary spots on the microarray.

Hybridization of sample genomic DNA molecules can be monitored by measuring the intensity of fluorescence at particular spots on the microarray. Differences in the amount of hybridization between the samples can be readily observed. If only one sample’s nucleic acids hybridize to a particular spot on the microarray, then that spot will appear either green or red. However, if both samples’ nucleic acids hybridize, then the spot will appear yellow due to the combination of the red and green dyes.

Although microarray technology allows for a holistic comparison between two samples in a short time, it requires sophisticated (and expensive) detection equipment and analysis software. Because of the expense, this technology is typically limited to research settings. Researchers have used microarray analysis to study how gene expression is affected in organisms that are infected by bacteria or viruses or subjected to certain chemical treatments.

a) A diagram of a microarray. First two different cells are grown in culture – in this case cancerous and healthy cells. Then RNA is isolated from each culture. Then reverse transciptase labeling produces cDNA with either a red fluorescent probe or a green fluorescent probe. In this case the cDNA from the cancerous cells is produced using red fluorescent probes and the cDNA from healthy cells is produced using green fluorescent probes. These are then hybridized onto a microarray – which looks like a glass slide with many etchings. B) The result on a computer compares all the genes that are active in the cells. Green regions indicate genes that are only expressed in heathy cells, red regions indicate genes that are only expressed in cancerous cells, yellow regions indicate genes that are expressed in both cancerous and healthy cells, and black regions indicate genes that are not expressed in either cell type.
(a) The steps in microarray analysis are illustrated. Here, gene expression patterns are compared between cancerous cells and healthy cells. (b) Microarray information can be expressed as a heat map. Genes are shown on the left side; different samples are shown across the bottom. Genes expressed only in cancer cells are shown in varying shades of red; genes expressed only in normal cells are shown in varying shades of green. Genes that are expressed in both cancerous and normal cells are shown in yellow.
  • What does a DNA probe consist of?
  • Why is a Southern blot used after gel electrophoresis of a DNA digest?

Questions & Answers

how does Neisseria cause meningitis
Nyibol Reply
what is microbiologist
Muhammad Reply
what is errata
Muhammad
is the branch of biology that deals with the study of microorganisms.
Ntefuni Reply
What is microbiology
Mercy Reply
studies of microbes
Louisiaste
when we takee the specimen which lumbar,spin,
Ziyad Reply
How bacteria create energy to survive?
Muhamad Reply
Bacteria doesn't produce energy they are dependent upon their substrate in case of lack of nutrients they are able to make spores which helps them to sustain in harsh environments
_Adnan
But not all bacteria make spores, l mean Eukaryotic cells have Mitochondria which acts as powerhouse for them, since bacteria don't have it, what is the substitution for it?
Muhamad
they make spores
Louisiaste
what is sporadic nd endemic, epidemic
Aminu Reply
the significance of food webs for disease transmission
Abreham
food webs brings about an infection as an individual depends on number of diseased foods or carriers dully.
Mark
explain assimilatory nitrate reduction
Esinniobiwa Reply
Assimilatory nitrate reduction is a process that occurs in some microorganisms, such as bacteria and archaea, in which nitrate (NO3-) is reduced to nitrite (NO2-), and then further reduced to ammonia (NH3).
Elkana
This process is called assimilatory nitrate reduction because the nitrogen that is produced is incorporated in the cells of microorganisms where it can be used in the synthesis of amino acids and other nitrogen products
Elkana
Examples of thermophilic organisms
Shu Reply
Give Examples of thermophilic organisms
Shu
advantages of normal Flora to the host
Micheal Reply
Prevent foreign microbes to the host
Abubakar
they provide healthier benefits to their hosts
ayesha
They are friends to host only when Host immune system is strong and become enemies when the host immune system is weakened . very bad relationship!
Mark
what is cell
faisal Reply
cell is the smallest unit of life
Fauziya
cell is the smallest unit of life
Akanni
ok
Innocent
cell is the structural and functional unit of life
Hasan
is the fundamental units of Life
Musa
what are emergency diseases
Micheal Reply
There are nothing like emergency disease but there are some common medical emergency which can occur simultaneously like Bleeding,heart attack,Breathing difficulties,severe pain heart stock.Hope you will get my point .Have a nice day ❣️
_Adnan
define infection ,prevention and control
Innocent
I think infection prevention and control is the avoidance of all things we do that gives out break of infections and promotion of health practices that promote life
Lubega
Heyy Lubega hussein where are u from?
_Adnan
en français
Adama
which site have a normal flora
ESTHER Reply
Many sites of the body have it Skin Nasal cavity Oral cavity Gastro intestinal tract
Safaa
skin
Asiina
skin,Oral,Nasal,GIt
Sadik
How can Commensal can Bacteria change into pathogen?
Sadik
How can Commensal Bacteria change into pathogen?
Sadik
all
Tesfaye
by fussion
Asiina
what are the advantages of normal Flora to the host
Micheal
what are the ways of control and prevention of nosocomial infection in the hospital
Micheal
what is inflammation
Shelly Reply
part of a tissue or an organ being wounded or bruised.
Wilfred
what term is used to name and classify microorganisms?
Micheal Reply
Binomial nomenclature
adeolu

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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