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Gel electrophoresis

Because nucleic acids are negatively charged ions at neutral or alkaline pH in an aqueous environment, they can be moved by an electric field. Gel electrophoresis is a technique used to separate charged molecules on the basis of size and charge. The nucleic acids can be separated as whole chromosomes or as fragments. The nucleic acids are loaded into a slot at one end of a gel matrix, an electric current is applied, and negatively charged molecules are pulled toward the opposite end of the gel (the end with the positive electrode). Smaller molecules move through the pores in the gel faster than larger molecules; this difference in the rate of migration separates the fragments on the basis of size. The nucleic acids in a gel matrix are invisible until they are stained with a compound that allows them to be seen, such as a dye. Distinct fragments of nucleic acids appear as bands at specific distances from the top of the gel (the negative electrode end) that are based on their size ( [link] ). A mixture of many fragments of varying sizes appear as a long smear, whereas uncut genomic DNA is usually too large to run through the gel and forms a single large band at the top of the gel.

Photo shows a black background with 9 faint gray vertical bands (lanes). In those bands are horizontal white slightly blurry bands of varying thicknesses and brightness. The faint gray lanes on the left and right edges have a lot of horizontal bands, and the 7 in the middle have only a few each, in different positions.
Shown are DNA fragments from six samples run on a gel, stained with a fluorescent dye and viewed under UV light. (credit: modification of work by James Jacob, Tompkins Cortland Community College)

Polymerase chain reaction

DNA analysis often requires focusing on one or more specific regions of the genome. It also frequently involves situations in which only one or a few copies of a DNA molecule are available for further analysis. These amounts are insufficient for most procedures, such as gel electrophoresis. Polymerase chain reaction (PCR) is a technique used to rapidly increase the number of copies of specific regions of DNA for further analyses ( [link] ). PCR uses a special form of DNA polymerase, the enzyme that replicates DNA, and other short nucleotide sequences called primers that base pair to a specific portion of the DNA being replicated. PCR is used for many purposes in laboratories. These include: 1) the identification of the owner of a DNA sample left at a crime scene; 2) paternity analysis; 3) the comparison of small amounts of ancient DNA with modern organisms; and 4) determining the sequence of nucleotides in a specific region.

Figure showing PCR in 4 steps. First, the double strand of DNA is denatured at 95 degrees Celsius to separate the strands. The 2 strands are then annealed at approximately 50 degrees Celsius using primers. DNA polymerase then extends the new strands at 72 degrees Celsius. The fourth step shows that this procedure takes place many times, resulting in an increase in copies of the original DNA.
Polymerase chain reaction, or PCR, is used to produce many copies of a specific sequence of DNA using a special form of DNA polymerase.

Cloning

In general, cloning    means the creation of a perfect replica. Typically, the word is used to describe the creation of a genetically identical copy. In biology, the re-creation of a whole organism is referred to as “reproductive cloning.” Long before attempts were made to clone an entire organism, researchers learned how to copy short stretches of DNA—a process that is referred to as molecular cloning.

Molecular cloning

Cloning allows for the creation of multiple copies of genes, expression of genes, and study of specific genes. To get the DNA fragment into a bacterial cell in a form that will be copied or expressed, the fragment is first inserted into a plasmid. A plasmid    (also called a vector in this context) is a small circular DNA molecule that replicates independently of the chromosomal DNA in bacteria. In cloning, the plasmid molecules can be used to provide a "vehicle" in which to insert a desired DNA fragment. Modified plasmids are usually reintroduced into a bacterial host for replication. As the bacteria divide, they copy their own DNA (including the plasmids). The inserted DNA fragment is copied along with the rest of the bacterial DNA. In a bacterial cell, the fragment of DNA from the human genome (or another organism that is being studied) is referred to as foreign DNA to differentiate it from the DNA of the bacterium (the host DNA).

Questions & Answers

difference between DNA and RNA
Eyitayo Reply
what determines the aeration level in the soil
Shola Reply
what is homeostasis?
Sarita Reply
What is biology
Don Reply
Biology z the study of life
GOLDEN
what's biology
buran
biology is the study of living nd none living organism
Chinaza
Biology is the study of life
Quadri
yes Sir
Said
what's cell biology
Prince
biology is the study of life
Raheal
what is asexual reproduction,?
Awoi Reply
A type of reproduction which does not involve the fusion of gametes or a change in the number of chromosomes
Serena
Reproduction without sex... In which form a single organism or cell makes a copy of itself.
Serena
Please explain the concept of mitosis and meiosis
Serena
I guess you could use it for study buddies and brushing up on what you need to
Serena
what is mitosis
Klp
Asexual reproduction?
Serena
why pepsin and trypsin released in active form?
Yacqub
mitosis is the type cell division in which two daughter cells have same no. of chormosomes
syed
Hi
Don
chromosome number remains the same in mitosis
mc
Ello
Henry
Hii
Nikky
Yrr help me.
Nikky
Physical chemistry..... Koi h jo mujhe physical chem ki notes send kr ske
Nikky
what is asexual reproduction
Targbe
what makes golgi body in plants
Abdulkareem Reply
name the membrane of the plants
Abdulkareem
how can turners syndrome be corrected before birth
Balinda
which animal survive from being preyed just because of being humble, slow, and not aggressive
Balinda
During organs transplantation, the organs cannot be taken from just anybody since the graft would be rejected sooner or later due to
Liter Reply
Non-MHC compatibility on the organ and an attack from the patient's immune system.
Eric
what makes golgi body in plants
Abdulkareem
why trypsin and pepsin released in active form
Yacqub
what is integument system
Joy Reply
Cellular respiration
Lucy Reply
what are the characteristics of living things
Ruth Reply
Movement Respiration Nutrition/Feeding Irritability/Sensitivity Growth Excretion Reproduction Deat/Life span
Hashim
What makes children from the same father and mother sometimes don't look alike?
Hashim
identification of problems
Nana Reply
what happens in the process of raising the human arms
Nana
what is biology
Brandi Reply
first step in scientific method
Brandi
In an investigation the pancreatic duct of a mammal was blocked.It was found that the blood sugar regulation remained normal while food digestion was impaired.Explain
Mac Reply
To begin with, obstruction of pancreatic duct will alter the blood sugar level as the juices responsible for glucose regulation will be rendered inconsequential. This will in turn affect the rate of digestion and absorbtion of digested food substances by the Villus .
Muktar
characteristics of algae
OMIME Reply
Algae are eukaryotic organisms. Algae do not have roots and stems. Algae have chlorophyll and helps in carrying out photosynthesis.
Aditi
Cell wall is the rigid layer enclosed by membranes of plants and prokayortic cell, it maintains the shape of the cell and serve as a protective barrier.
chizoba Reply
ECOLOGY: is a branch of biology that studies the interactions among organisms and their biophysical environment, which includes both biotic and abiotic components. 
chizoba
via nutrient cycles and energy flows. For instance, the energy from the sun is captured by plants through photosynthesis. Photosynthesis is a biological process through which plants manufacture their own food with the aid of light from the sun and frc sources (e.g. cabon dioxide and water)
chizoba

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Source:  OpenStax, Concepts of biology. OpenStax CNX. Feb 29, 2016 Download for free at http://cnx.org/content/col11487/1.9
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