Northern blot technique Of RNA - Principle , Steps | Biology Blog

                                          Northern blotting 

Hello guys! In this blog, we are going to explore the northern blot technique. This technique can be used in any viral detection test as the genetic material of the virus is RNA.

So, before moving to northern blot we focus on the word - blot.

Blot is basically a technique in molecular biology and genetics in which we DNA/RNA or proteins could be transferred onto the carrier membrane. The nylon membrane or nitrocellulose membrane are used for blotting purpose. 

The transferring of molecules like DNA/ RNA or proteins to the membrane can take place after gel electrophoresis. After blotting the transferred molecules are visualized. 

For visualization, we can use the following methods- 

1. If the probe is labeled with the radioactive chemical (32P), it can be observed on X-ray film directly. 

2. If a chromogenic detection is used, it can be visualized by the development of color on the membrane itself. 

3. For the detection of DNA/RNA, a labeled probe is used. The labeling can be done with the radioactive isotopes (32P) or fluorescent dyes. 

So this is the blot technique, now we concentrate on the main topic - what is Northern blot and what is its steps.

Northern blot technique -  so guys, this technique was developed by Alwine and coworkers in 1977 and modified by  Thomas in 1980. In this technique, we can detect the presence of particular RNA in tissue samples.

Steps - it has the following steps - 

1. Isolation and purification - The RNA of interest is isolated by a high-speed centrifugation method and then it needs to purify.

2. Denaturation and electrophoresis - Denaturation is done for removing the fold in RNA and for this formaldehyde are used. After denaturation the shape of RNA becomes linear. Now, RNA is mixed with buffer solution and formaldehyde and run through gel electrophoresis. The electrophoresis separate RNA fragments. 

3. Blotting - After electrophoresis, blotting us done for transferring RNA into the nylon membrane. 

4.Incubation with the labeled probe - The membrane containing RNA is now incubated with a labeled probe. The probe used should be complementary to the RNA used. The probe used may be labeled with a radioactive chemical, not a fluorescent dye.

5. Hybridization- The membrane blot is soaked in polyethylene glycol and then washed in solution to remove unbound or loosely bound probe. 

6. Detection-  The labeled can be visualized by autoradiography. 

 Have you read - Western blotting techniques

convolascent plasma therapy for cornonavirus

Hello guys!

 The elevating cases of corona in the world forced researchers to discover antibiotics.  scientists experimenting and researching continuously to develop vaccines or therapy that may help to beat the corona crisis.

So, one of the therapies that doctors believe to experiment on patients is plasma therapy. Plasma is a watery fluid of the blood. This plasma is consists of corpuscles and thrombocytes in addition to that plasma also contains some proteins (immunoglobulin) which produce antibodies.
Our body works on the principle of the immune system. The basic principle of the immune system is that it forms antibodies against any foreign antigen that enters the body. Our immune system memorizes this antigen and this is called the primary response. If the second time again this same antigen comes into the body by any means then our antibody recognizes it and neutralizes this foreign antigen.  This principle is becoming helpful for experimenting against coronavirus.

If any person recovered from this virus then the plasma of that person can be used to fight against this virus. These antibodies can be used to treat more patients who are suffering from this virus. The antibody memorizes this foreign antigen and neutralizes the virus. In this way, we could save multiple person' life suffered from covid-19.

Related Links  - Things to know about coronavirus 

                           Impact of coronavirus on mental health 

    

Past history of this therapy - 

• This therapy is recommended by the WHO in treating the Ebola virus.
• In 2015, this therapy is used in treating middle East respiratory disease which is also caused due to coronavirus
• At the time of the Spanish flu, this therapy was used experimentally.
• At the time of SARS, this therapy showed a good effect. 

Plasma therapy in present - Convalescent plasma therapy is explored by China and try it on some patients and they revealed that the patient showing improvement in recovery was noticed and all of them later on discharged. This exploration was published by China in the Journal of American Medical Association. 

However, this process in under trial as the research utters that there is some unknown right that needs to resolve out like in what quantity doses will work as antibodies? At what stage of illness it will be given to the patient? Are some reasons that will need to be addressed.

Things to know about pandemic disease coronavirus : Biology Blog

                                        Coronavirus

Coronavirus is an infectious disease that is spreading rapidly all over the world. More than 20 lakh people of the world have been affected by this virus and incoming time its chances of increase will be more. It is coming from the Wuhan, the capital of China on 29 December 2019 and since then it is spreading more fastly.

Symptoms - Fever, cold, cough, fatigue, diarrhea are general symptoms of this virus.  Older people and the child under 10 years of age are prone to this pandemic disease. The symptoms of this disease will appear between two to fourteen days. The people having a pre-existing disease like cancer, respiratory problems and diabetes have more risk to get the infection.

Mode of transmission - This virus is inactive until they do not get any host body to replicate their genome. After entering into the host body they first attach to the respiratory system and block the system, a consequence of that person feel some cough problem. The virus lives here for some days and multiplies their copy and after that slowly affects the whole body and then the person feels muscular pain and then fever is resulted out.

Its mode of transmission is just similar to the normal flu. If any normal person comes in contact with the affected one then this virus jumps into that person and in results, the person develops symptoms. 

Similarly, if the person touches the contaminated surface and then touches their mouth, nose, and ear then this virus gets its way to enter into mouth. This virus is active on the surface up to 72 hrs.
Have you read- Impact of coronavirus on mental health  of people 

Preventive measures to adopt - 

• Always use the mask to cover your mouth and nose area.
• Avoid close contact with the affected person.
• If for any reason or unknowingly you come in contact with the person having symptoms then isolate yourself for a minimum of 14 days.
• Follow social distancing.
• Stay at home.
• Regular drink warm water at least once in a day.
• Wash your hands with soaps at least 20 times a day.
• Use sanitizer.

Explain transgenic animals and ethical issues : Biology Blog

                             Transgenic Animals & ethical issues 

Hello guys!

on this blog, we are going to discuss what are the transgenic animals and what are the issues related to that. So, guys, transgenic animals are those animals that have their DNA  manipulated so that the extra foreign gene can be expressed. There are multiple examples of transgenic animals like sheep, cows, mice, rats, etc.

Reasons for developing transgenic animals -  

• To the understanding of gene regulation and their effect on normal physiology of the animal and its development. For example, we can study various growth factors.
• The study of the disease can be done by studying genes, which are responsible for causing disease in human and their treatments.
• Biological products can be produced by introducing the portion of DNA, which codes for a particular product into transgenic animals. For example - human protein ( alpha-antitrypsin ) is used to treat emphysema.
• Rosie, the first transgenic cow was produced in 1997 for the purpose of producing protein-enriched milk. This milk contain the human alpha - lactoalbumin and was nutritionally balanced for human babies.
  
• For testing vaccine.
• For testing chemical safety transgenic animals are required.

Method representation -

Ethical issues -

• When this type of organism is exposed to the environment , there are chances of unpredictable results.
• The modification/usage of living organisms for public service has also created problems with patents granted for the same.
• In order to control these issues. the Indian government has set up an organisation such as GEAC, which makes a decision regarding the validity of genetically modified research and the safety of introducing GM organisms for public services.  

Why Living Organisms Are Classified | Biology Blog

                                        Living Organisms 

Hello Guys!


We'll talk about why it's important to categorize living things. Therefore, before moving on to the subject, let's first talk about something really fundamental: What is Living?

 

When we consider what it means to be alive, our minds automatically focus on characteristics like growth, metabolism, the ability to self-replicate, etc. 

Let's talk about each of them:

Growth -  Growth refers to an abrupt rise in size and quantity. Similar to how our fingers reach a specific height before stopping to develop, our height also rises for a limited time before stopping, however in plants, growth continues continually for the duration of their lives. Cell division is also how unicellular creature grow.  But as I mentioned earlier, growth is another name for a gain in mass. Using these standards, we can also assert that nonliving things, such as the collection of objects on any surface, mountains, etc., grow.  Consequently, the result is that internal growth is a trait of living organisms and occurs when they are alive.

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Reproduction - Reproduction is the biological process by which a parent can create kids of same species. Therefore, this is another trait of a living being. There are two different types of reproduction processes: asexual reproduction and sexual reproduction.  The reverse of sexual reproduction is asexual reproduction, in which only one parent is responsible for producing their own sort of children.

          

        However, there is one exception to this rule: not all organisms are capable of reproducing, i.e., some couples are unable to conceive a child as a result of infertility. Therefore, we cannot conclude that reproduction is a characteristic shared by all living things.

Metabolism - Chemical compounds make up every living thing. The size of the compounds can vary, and they can ultimately transform into specific biomolecules. Chemical reactions are used to describe this change. Basically, our bodies go through a huge number of chemical reactions, and the aggregate of all of these is what is known as metabolism. Catabolism is the process by which complex molecules undergo chemical reactions to become simpler ones, while anabolism is the process by which a simpler substance becomes complex, as in the case of our muscular system. Such a characteristic is unique to living things because it is absent from nonliving things.

 You can also read it in -Biology and features of living organisms 

The need for Classification - On earth, there are many different kinds of bacteria, animals, and plants. The size, form, color, habitat, and many other characteristics of all these species vary. As a result, studying each of them is impossible. As a result, scientists have developed a system to categorize all living things. These methods of classification are based on rules and principles that allow identification, nomenclature and finally classification of organisms. 

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Why Living Organisms Are Classified

Hemophilia - A Genetic Disorder : Biology Blog

                                        Hemophilia

With haemophilia, a hereditary condition, blood continues to ooze after a minor injury, a seizure, or even after you pull out your tooth. This happens as a result of the body's lack of a blood clotting factor. Internal bleeding that affects the body's major organs, such as the brain, muscles, or joints, can occasionally occur. This type of bleeding is more serious.

When we are harmed, our blood typically oozes out for a while before coagulation forms to prevent more bleeding. This is made possible by the body's 8th and 9th blood clotting factors, which regulate bleeding. If these factors are missing or absent, this condition develops.

Symptoms- The symptoms of this disease may vary depending upon the level of the clotting factor.

    Types Of hemophilia - There are two types of hemophilia - hemophilia A and hemophilia B.

        The gene F8, which also gives instructions for producing blood clotting factor 8, is absent or mutated in haemophilia A, which leads to the disease. Haemophilia B is brought on by mutations in gene 9 that provide instructions for producing blood clotting factor 9th later ( also known as Christmas factor ). Together, these two genes support the blood clotting process.



Inheritance pattern - The problematic gene is frequently found on the X chromosomes in common situations. We are aware that each person inherits one set of sex chromosomes from their father and mother. Males inherit one X and one Y chromosome from their parents, respectively. The mother gave each female one chromosome, while the father gave each female an X chromosome.

This indicates that this illness only affects men and is always passed from mother to son. Women act as the disease's carriers.

Risk Factor - family history is the biggest risk factor for the inheritance of this disease.

Acquired Haemophilia - Although 30% of people with haemophilia do not have a family history of the condition, the ailment typically develops as a result of pattern inheritance.

A uncommon illness called acquired haemophilia causes the body's autoimmune system to become active and kill the cells that produce blood clotting factors. It may be related to multiple sclerosis, cancer, or pregnancy.

  

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How HIV Virus Replicate To Cause Immunodeficiency: Biology Blog

                                               HIV/ AIDS 

HIV virus is generally a human Immunodeficiency virus that is the dangerous virus that attacks the immune system of the human and affects the fighter cells of the body (lymphocytes cells). The person suffered from this disease is more vulnerable to sickness. This disease is dangerous because someone can be unknowingly transferred to the other and the symptom of the disease appear after so many times. Although there are some medications available for inhibiting the spreads and lifespan of the sick person however still there is no cure for HIV.

Modes Of Transmission -  having Sexual Intercourse with more than one person.
                                            Sharing blood/blood products
                                            Sharing the same needle
                                            From infected mother to the child   


Symptoms -

 Replication Cycle -   

• The HIV virus attacks the macrophages cells in the human body.    
• RNA(the genetic material of virus) is replicated to form viral DNA by the enzyme reverse transcriptase.                                                                                                                           
  
• Viral DNA  now gets incorporated into the host cell's DNA and directs the infected cells to produce viruses.
• Macrophages continue to produce virus particles and function as HIV factories.
• The virus particles enter helper T-lymphocytes in the blood, where they continue to replicate and produce viral progenies.
• The number of T-lymphocytes progressively decreases in the body of the infected person.
• With the decrease in the number of T-cells, the immunity also decreases. The person is unable to produce an immune response even against common bacteria like mycobacteria, parasites like toxoplasma, viruses, and fungi.  

Test -   ELISA test is conducted for the confirmation of the disease in which the blood sample of the sick person is taken and analyzed. The full form of ELISA is the enzyme-linked immunosorbent assay.

Prevention Measure for HIV -   There are certain measures by which we can prevent people from this virus-like,  With the help of various NGOs educating the people about the disease, ensuring the use of disposable syringe or needles,   by free distribution of condoms, etc.