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What is gel electrophoresis | Types and principles

What is gel electrophoresis | Types and principles

Gel Electrophoresis is a widely used analytical method in molecular biology and biotechnology.

Widely practiced for the separation of particles like viruses, small DNA fragments and nucleic acids and proteins. Also used after the PCR, to separate the DNA copies according to their quality.

Moreover, the electrophoresis method relies on the basic principle that the charged particles in a sample will migrate if an electric field applied to it.

Therefore, once electric field applied, the particle in the solution will migrate according to the charge or size.

Above all, widely used gels like polyacrylamide and agarose are commonly used .

This gel gives sieving or separating effect, and the particle separated by both size and charge.

Small molecules can travel faster than the larger one, also the negative charged particles will move towards anode. Similarly, positive charged particles will move towards cathode, in this way particle gets separated by size and charge.

Electrophoresis of cationic particle is called cataphoresis. Likewise, that of anionic particle called anaphoresis.

what is gel electrophoresis

Types of Electrophoresis

There are different types of electrophoresis but the most commonly and vividly used are mentioned below:

Capillary electrophoresis – molecules separated on the electrophoresis basis by using a capillary tube. To clarify, the molecules separated by running them through the capillary tubes. Accuracy higher but less quantity samples can be loaded at a time.

Native gels electrophoresis – protein run without SDS. Therefore, no proteins will get denatured in this method.

SDS – PAGE – Full form : Sodium Dodecyl Sulfate – Polyacrylamide gel electrophoresis. To clarify, SDS a detergent which can denature the protein and binds to the hydrophobic site of the molecule. Therefore, becoming the most dominant negative charge of the complex.

Here, the proteins are run through gel, polyacrylamide. Therefore, the particle density and size can be varied.

Electrofocusing gel – A gel with a pH gradient used here. As the particle travels through the gradient.

Meanwhile, it will loose protons and finally gets stucked at a point with no pH.

DNA Agarose gels – Here, agarose gels used to separate large fragments of DNA. The method used to separate according to the particle sizes. Here, no SDS needed because, DNA being negatively charged.

Above all, particle size sensitivity can be adjusted by adjusting gel density.

Sequencing gels – method used to separate smaller DNA fragments with a greater resolution. Firstly, the DNA denatured using heat and at the same time passed through polyacrylamide gel. Moreover, DNA which differ by 1 or 2 base can be separated more accurately.

The glowing DNA under the fluorescent light

Gel Electrophoresis applications

Applied for the estimation of DNA molecules.

Helps in mapping cloned DNA.

Analyzing the PCR products.

Gel electrophoresis widely used in forensics, molecular biology and genetic engineering.

Used in verification of amplified DNA using PCR.

Checking the quality of extracted DNA.

Gel electrophoresis a widely used technique, more modernized. Moreover, to determine the DNA fragment quality. In addition, used after the PCR to seperate and extract the DNA products.

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What is RT PCR? |Reverse Transcriptase PCR

What is RT PCR? |Reverse Transcriptase PCR
What is RT PCR

RT PCR or Reverse Transcriptase PCR is a test used to detect retroviral diseases like AIDS. It gained popularity during coronavirus outbreak.

Before knowing about RT PCR, please refer PCR or Polymerase chain reaction first here – https://sciencebirdie.com/2020/06/17/what-is-pcr-polymerase-chain-reaction-explained/

PCR or polymerase chain reaction is the technique in which the DNA is amplified and the billion DNA copies are made. But, it has exceptions like it can only detect only DNA viruses. Therefore, detection of RNA viruses like retroviruses cannot be possible.

What is RT PCR

This is where the RT PCR has the significance.

Working mechanism

Viruses like Coronavirus have RNA as the genetic material, therefore, the detection using the traditional PCR becomes impossible.

So, scientists convert RNA to DNA using an enzyme called Reverse transcriptase, the process known as Reverse Transcription. On the other hand, the conversion of DNA to RNA is called Transcription.

What is RT PCR
Thermocycler or PCR machine used in PCR technique

Steps in RT PCR

Firstly, the RNA converted to DNA using reverse transcription.

The DNA then transferred to the thermocycler machine or the PCR machine.

Afterwards, the steps denaturation, annealing and extension followed and the billion DNA copies extracted.

Please refer the PCR steps here – https://sciencebirdie.com/2020/06/17/what-is-pcr-polymerase-chain-reaction-explained/

Advantages

Coming to advantages, it had got more in addition than the traditional PCR.

Highly sensitive as the RNA involvement in it.

Highly specific, with more accuracy than PCR.

Requires less than 4 hours in diagnosis, moreover, the laboratories take up to 6-8 hours.

As the genetic material placed in a closed tube with enzymes, it has low potential contamination.

RT PCR in Corona test

Nowadays, with Coronavirus being prevalent, the RT PCR use got very much boosted up.

What is RT PCR

Firstly, the victim’s blood sample is taken then using several enzymes the RNA is extracted and converted to DNA and with PCR many copies are made.

Now, the DNA bases are made to pair with another bases tagged with markers. Further, the pairs are examined through fluorescence technique. If there are abnormalities, in the pairs the pairing does not occur. In other words, the person stated positive in the test.

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TARDIGRADES

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Hi! The water bear - the Arnold Schwarzenegger of the micro bug world - raises a smile to the camera

INDEX

Tardigrades are found everywhere. They are present within 100 meters of a person. They are indestructible. Importantly they are also called moss piglets or water bear.

Initially they are 1 mm in size, their length is 1.5 mm after growth. . They are aquatic invertebrates.

CLASSIFICATION

The Tardigrades belong to the kingdom Animalia. It is placed in Tardigrada phylum.

About 1150 species of tardigrades are found. Apart from this they are also known as slow stepperss.

DEATH OF TARDIGRADES

They are one of the immortal creatures. According to scientists, they can also survive an asteroid impact.

They can live in adverse conditions. They can live under high pressure. Also in high temperature and low temperature conditions.

They can live up to 30 years without food. They can also live in and near volcanoes.

Eventually, the ultimate death of the sun can erase them.

EXPERIMENTS

In 2008, the European Space Agency (ESA) sent the Tardigrades into space. They managed to avoid cosmic rays and rather harmful UV rays which are 1000 times more powerful than Earth. They can survive to extremes conditions in vacuum. Also they are the first creatures. to do this.In addition, their DNA is used for advanced studies.

A type of protein found in them named Dsup, which protects DNA and cells from extreme conditions and desiccation. This gives rise to further advanced studies and experiments.

This DNA can be induced in humans.

As a result helping to protect humans from harsh conditions. People working in nuclear plants will get benefits to prevent themselves from radiation.

MEDICAL FIELD

One study found that cells involving a human and a tardigrade cells were able to prevent up to 40% damage by X-rays.

It opens up new ways of treating cancer. People undergoing radiation treatment will have new expectations because prevention will be easier.

Cosmic rays and UV rays therefore it will provide more effective preventive methods for astronauts.

IMMORTAL ANIMALS

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Immortal animals found in earth

JELLY FISH(SEA JELLIES)

~ jellyfish ~

Organisms related to the phylum Cnidaria. They are found in deep oceans. Actually considered as immortal animal. Above all,they are mostly without brain and blood.

Almost every species is invertebrate. They can reproduce both sexually and asexually. Immortal animals like Territopsis dohrnii can start a new life cycle and is considered immortal.

Above all Jelly fish contain stem cells that can regenerate or renew cells.

PLANARIAN FLATWORM

The #planarian #flatworm doing its amazing dividing trick.

Immortal animals.In other words,they are flat worms belonging to the phylum platyhelminths. mostly carnivorous.

The experiment showed that the dissection of a flatworm from either of the two sides would produce two identical worms .

The whole body of two parts takes different ways. Pluripotent stem cells allow this.Here’s the video – https://www.youtube.com/watch?v=hTC1eNTBXvE

LOBSTERS

Maine Lobster – Santa Barbara Fish Market

Immortal animals are very lucky, but eaten as seafood. They are truly immortal until fishermen catch them. related to the Phylum arthropoda.

An enzyme telomerase is present. It prevents DNA damage and helps in replication and renewal. .No built in life expectancy. Therefore,the cells are constantly renewed.

HYDRA

Louis Shalako: Hydra.

Considered immortal.These organisms have regenerative potential. They can breed in two ways. In other words sexually and asexually.

Hydra (plural for hydra) who die sexually and those who breed asexually remain immortal.

Also most of them reproduce by the budding which is actually asexual.

TARDIGRADES

One of my favorite creatures. These cute little ones are 1 mm in size. Above all, they can survive harsh temperatures, pressures and radiation. .

In conclusion death of a star can kill Tardigrades. Actually considered immortal animals.

In addition, a protein called Dsup is present in them. It helps in surviving harsh conditions

SUPERBUGS

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Walmart Pork Found To Contain Deadly “SuperBugs” Resistant To Antibiotics - YouTube

What are Superbugs?

Superbugs are the bacteria which have became resistant to antibiotics.

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The use of antibiotic had increased so rapidly that it is being used for normal fevers,while it shouldn’t be used.

Therefore, the bacterias are developing resistance and becoming superbugs,in addition, the resistance capability of our immunity is decreasing.

Countries where the use of antibiotics are controlled like Australia,Germany etc,the growth is far more less.

But in countries like India,Greece etc, where the use of antibiotic is rapid the growth of antibiotic growth and rise is rapid.

PREVENTION

It is always better to be preventive from any infection so:

  • Follow proper hygiene to prevent any infection or disease
  • Do not use antibiotic for normal fevers
  • Similarly,do not use antibiotic for viral infections
  • Above all, boost your immune system by regular exercise and consume healthy foods.

SYMPTOMS

No symptoms especially for superbug because the infections are always confused with the infection of normal bacteria or pathogens.

Superbugs are diagnosed after prolonged treatment against the bacteria. If no changes then it is a superbug.

Antibiotic resistant superbugs killing thousands: What to know

SUPERBUGS IN INDIA

In India the overuse of antibiotic had caused great increase of the Superbug.The hospitals are the main breeding grounds for the superbug.Consequently,newborns suffer more likely from the Superbugs.

 In addition, poor sanitation also play a role in growth of these bacterias.

SUMMARY

Superbugs are the bacteria which have became resistant to antibiotics .

Prevention is better in case of Superbug infection.Superbug infection is difficult to treat.

Reduce antibiotic use .

Doctors should stop encouraging the overuse of antibiotics