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Forensic scientists use RFLP analysis as a form of DNA fingerprinting , which is useful for analyzing DNA obtained from crime scenes, suspects, and victims. DNA samples are collected, the numbers of copies of the sample DNA molecules are increased using PCR , and then subjected to restriction enzyme digestion and agarose gel electrophoresis to generate specific banding patterns. By comparing the banding patterns of samples collected from the crime scene against those collected from suspects or victims, investigators can definitively determine whether DNA evidence collected at the scene was left behind by suspects or victims.

A diagram of RFLP analysis. Two strands of DNA are shown; a “normal” strand and a mutated strand. The normal strand has 3 locations where the restriction enzyme MSTI cuts. The mutation on the other strand destroys one of the restriction sites. As a result, the mutated strand has one fewer bands on a gel.
RFLP analysis can be used to differentiate DNA sequences. In this example, a normal chromosome is digested into two fragments, whereas digestion of a mutated chromosome produces only one fragment. The small red arrows pointing to the two different chromosome segments show the locations of the restriction enzyme recognition sites. After digestion and agarose gel electrophoresis, the banding patterns reflect the change by showing the loss of two shorter bands and the gain of a longer band. (credit: modification of work by National Center for Biotechnology Information)

Southern blots and modifications

Several molecular techniques capitalize on sequence complementarity and hybridization between nucleic acids of a sample and DNA probes. Typically, probing nucleic-acid samples within a gel is unsuccessful because as the DNA probe soaks into a gel, the sample nucleic acids within the gel diffuse out. Thus, blotting techniques are commonly used to transfer nucleic acids to a thin, positively charged membrane made of nitrocellulose or nylon. In the Southern blot technique, developed by Sir Edwin Southern in 1975, DNA fragments within a sample are first separated by agarose gel electrophoresis and then transferred to a membrane through capillary action ( [link] ). The DNA fragments that bind to the surface of the membrane are then exposed to a specific single-stranded DNA probe labeled with a radioactive or fluorescent molecular beacon to aid in detection. Southern blots may be used to detect the presence of certain DNA sequences in a given DNA sample. Once the target DNA within the membrane is visualized, researchers can cut out the portion of the membrane containing the fragment to recover the DNA fragment of interest.

A diagram of a Southern blot. First, electrophoresis is used to separate DNA fragments by size. There can be so many fragments that they appear as a smear on the gel. Next DNA is transferred from the agarose gel to a nylon membrane. The buffer moves from the wet sponge below up through the gel and the membrane into the paper towels. It carries the DNA fragments with it until they become caught in the membrane. Finally, the membrane is based in a solution containing a probe, a short piece of DNA complementary to the sequence of interest. The probe is labeled or tagged with a fluorescent dye so that the location of DNA fragments to which it hybridizes can be visualized.
In the Southern blot technique, DNA fragments are first separated by agarose gel electrophoresis, then transferred by capillary action to a nylon membrane, which is then soaked with a DNA probe tagged with a molecular beacon for easy visualization.

Variations of the Southern blot—the dot blot, slot blot, and the spot blot—do not involve electrophoresis, but instead concentrate DNA from a sample into a small location on a membrane. After hybridization with a DNA probe, the signal intensity detected is measured, allowing the researcher to estimate the amount of target DNA present within the sample.

A colony blot is another variation of the Southern blot in which colonies representing different clones in a genomic library are transferred to a membrane by pressing the membrane onto the culture plate. The cells on the membrane are lysed and the membrane can then be probed to determine which colonies within a genomic library harbor the target gene. Because the colonies on the plate are still growing, the cells of interest can be isolated from the plate.

Questions & Answers

what is prokaryotes
PRALHAD Reply
are micro-organisms that have no nucleus e.g bacteria
Foster
and they do not possess membrane bound organelles e.g mitochondria
Bharat
yes
Foster
they are single cellular organisms,
Rajasekhar
all the organelles are embedded in cytoplasm of the cell
Rajasekhar
They don't possess true nucleus
PREMI
they're unicellular organisms
Kamaluddeen
they are classified as bacteria and. archaea. bacteria have a cell membrane conposed of peptidoglycan and archaea is composed of psuedomu
Tara
pseudomurein
Tara
organelles are non membraneous as the nucleus
Tara
What is the biosynthesis of a bacteria cell
Adesina Reply
biosynthesis is multiple step enzymes catalyze process which occur in bacterial cell
Jayeeta
biological way of manufacturing food and utilizes it for growth, multiplication and development
Kamaluddeen
prokaryot
PRALHAD
what is microbiology
Kiirya Reply
which organism can not be seen necked eyes is knowm as microbiology
meet
microorganisms
Jayeeta
micro can be define as d smallest living organisms dat can not be see wit naked eyes only with the aid of a microscope 🔬
muhammed
bio means life logy means study
muhammed
biology can b define as the systematic study of living things
muhammed
microbiology can be define as the study of tiny organisms, that's according to my understanding
muhammed
Microbiology is the study of living organisms which cannot be seen by the naked eye but only with the aid of a microscope. These microorganisms can be classified as bacteria, fungi, micro-algae, protozoa and virus.
Tatz
microbes that can't seen in naked eyes. upto 100micrometer we can see in naked eyes but 1 microns or nanometers are invisible in naked eye.
Jayeeta
what is a cell
Oteri Reply
cell can be define as smallest indivisible organism in the body
muhammed
cell is the smallest unit which makes up the whole body
Yue
cell is smallest unit and there are two types prokaryotic cell and eukaryote cell. bacteria a prokaryotic cell which works independently.
Jayeeta
nice forum
muhammed
Which parasite competes with the host for vitamin B12 to cause pernicious anaemia
Abubakar
Diphyllobothrium latum for vit B12
Jayeeta
it is also fish tapeworm
Jayeeta
Diphyllobothrium sp feed vitB 12
Jayeeta
recent trends in microbiology of biofertilizer
Ykomalipriya Reply
using nano technology
Jayeeta
what is transposors
EMMANUEL Reply
Junk DNA, jumping gene.
Pradeep
transposon elements are jumping gene as DNA sequence move from one location to anothet
Jayeeta
Yes
Pradeep
what is microbiology
Usama Reply
Microbiology is the study of organisms, most of which are too small to be seen with the naked eye, including bacteria, algae, protozoa, and fungi, as well as acellular agents, such as viruses and prions.
digvijaya
what is gram +ve and gram - ve bacteria?
Sharma
gm- contain lipid layer so it take counter stain during gram staining and pink or red in color but gm + does have thick LPS layer.
Jayeeta
what year microbiology started
Kartik
Robert Koch was the first to detect microorganisms
Jayeeta
what appears microorganisms
Kartik
clear the questions please
Jayeeta
was it robert koch or robert hooke?
digvijaya
microbiology word came from microns. study microbes that not recognize in naked eyes.
Jayeeta
what are the branch of microbiology
Oyemade Reply
We can basically classify into pure and applied microbiology
Ozzie
what is arabinose operon and it's stucture
Pooja Reply
The L-arabinose operon, also called the ara or araBAD operon, is an operon required for the breakdown of the five-carbon sugar, L-arabinose, in Escherichia coli
Tharra
kya hum operon ko divide kar skate hain
Kartik
reactions if viruses to physical and chemical agents?
VICTOR Reply
All viruses contain the following two components: 1) a nucleic acid genome and 2) a protein capsid that covers the genome. Together this is called the nucleocapsid. In addition, many animal viruses contain a 3) lipid envelope. The entire intact virus is called the virion.
Asali
assume you have performed a gram stain on a sample of pus from a patients urethra. you see red, nucleated cells and purple rods. what can you conclude
Petty Reply
bacteria
ABUBAKAR
gram negative bacteria hence gonococcal infection
Kiirya
Study of micro organisation which we cannot see with naked eye...
shyam Reply
Despite their bad reputation, microbes are mostly beneficial or have a neutral effect on our lives. Microbiology is the scientific study of these microorganisms. Microorganisms are those organisms that are too small to see with the naked eye and include things like bacteria, fungi, and viruses.
Asali
yes
Dhirender
not 'things'...'organisms' like bacterias, viruses and fungi
Sinmi
simi, it's not bacterias but bacteria which is the plural form of the word while bacterium is the singular of it
Richard
bacterium is obvio in a singular form but plural of bacterium is bacterias
Sinmi
Richard is right it's bacterium which is the singular form and plural is bacteria. *Bacterias* doesn't exist.
Aakriti
that is microbiology which is deals with microbes only their structure
Kiirya
what is microbiology?
Sandeep Reply
what is microbiology
Glory
is the study of small microorganisms that can't be viewed by our naked eyes under a microscope
Nabukwasi
it's the study of microbes their classification, diversity, their cellular structure and processes, their applications and interaction with other organisms.
Chaitanya
what is EMP pathway
rajesh
which organism is implicated in meningoencephalitis
Peter Reply
Herpes virus
Carlos
Yeah
Laxmi
which mold is called dairy mold?
Gopinath
mushroomm
Top
Geotrichum candidum
Gopinath
hi
bio
Hlw
Laxmi
which infectious agent is the smallest
Sinmi
spore or virus
Anushree
prion ....it is a single protein in a misfolded form..there is also a theory based on it
Sinmi
yup...m sry
Anushree
yes it can be turned as a virus
Sinmi
which gram positive bacteria is responsible for food poisoning?
Sinmi
prion is not a living organism yet it is infectoois why
Jeevan
prions are virus like organism. and it is a misfolded protein that may cause disease to us.
Tharra
Clostridium perfringens, B.cereus
Sneha
Who is the father of the biology
Dhirender
I have one more question. . that who is the father of botany and medicine? ? can any one tell
Dhirender
define polar bond in simple way
Rashmi Reply
when there is formation of positive and negative poles i.e. one atom is +ve and other is -ve which further results in the dipole moment. Such bond is know as polar bond. It is a convalent bond only.
Aakriti

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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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