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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

must all prokaryotic cells posses a cell wall?
chris Reply
what is biology?
Cathy Reply
biology is basically the study of life
Robert
What is a celiac disease
Falase Reply
distinguish between properties and characteristics
Elee Reply
what are organelles
Elee
organelles are substances that makes up a cell
chris
what is biology
Prevail Reply
biology is science that studies life
Elee
it's the study of living and non living things
now that we have an estimate for the diameter of the cell.what estimate can we make about the volume of the cell?
faxhood Reply
why too much insulin result in low blood sugar
Leri Reply
For example, too much exercise can cost you to lose to much weight. Too much insulin will pull to much sugar out of your systemic system into your cells.
Eric
difference between DNA and RNA
Eyitayo Reply
DNA is deoxyribonuclaic acid. Deoxy refers to a lack of oxygen. The Ribose moity is missing an OH group. I think it is missing from the second C of the ring. RNA is ribonucleic acid. DNA has our genetic code in on it. RNA is translated from DNA and carries the blue print for protein synthesis.
Eric
The OH group on RNA prevents it from being reactive. But it is very unstable though. Would you want such a power tool floating around in your body, no. And you have three types of RNA: mRNA, tRNA and rRNA. Please let me know it this helped?😄
Eric
o
Browse
Eric answered this question perfectly
what determines the aeration level in the soil
Shola Reply
what is homeostasis?
Sarita Reply
It means balance in a biological system.
Eric
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
Biology is a science subject that deals with the study of living things and how they interact with there environment
chris
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
Plants have golgi body's also. Plants are eukaryotic cells. And membrane bound organelles are a characteristic of eukaryotic cells. Moreover golgi body's are creatted from the ER. Also do not forget plants have plastids and animal cells do not.
Eric
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
Let us remember MHC'S on our cells. This is how our cells determine self from n o n s e l f. Transplanted tissue has to have a certain amount markers. These have to match to the recipiant's markers. Even with this, immunosuppresant medacine is prescribed to the recipient.
Eric
Even with these measures the body may still reject the transport. This can occur even after the recipient excepting the transport for some time.
Eric
what is integument system
Joy Reply
This system is our skin. This includes the skin lining our alimentary system which includes the tissue from our mouth to our anus. Our skin is our largest organ system. It is mostly made up of epithelial tissue.
Eric

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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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