The average Researcher will spend about 4-5 years of time in the lab. The whole period is focused on setting goals and defining experiments, carrying out experiments, taking notes and analyzing results from experiments in the process of interpreting and concluding results of the experiment for publication of the outcomes. The most crucial steps is performing experiments that can be practical unlike other experiments that are theoretical and can be carried out. To get the most effective results from the tests it is vital to be proficient with the instruments as well as to be proficient with wet-lab methods.
Life science researchers are at the option of choosing any research area depending on their passions in the many research fields, including the molecular and cell biology biochemistry, genetics, and biotechnology for animals or plants. Each area of study is unique and has its own set of research projects that require different techniques and the use of various laboratory instruments. However, regardless of the words you’d like to write you have to learn the alphabet first. in a same way every researcher must master helpful lab techniques for success in wet-lab tests.
In this article, we’ll take a a review the lab Techniques that every researcher must know
1. Methods for Extraction and Storage of Biomolecules
Lab Techniques You Must Know The most frequently used biomolecules used in lab experiments are DNA and RNA. Protein. There are standard procedures to extract these biomolecules. The sensitivity of RNA to degradation must be extracted with extreme diligence. The extraction process involves the use of a variety of agents that play different roles in extracting the most pure biomolecule. It is essential for the researcher to understand the importance of each reagent, and then use it in a way that meets the specifications of the experiment. It is important to also be aware of a precise understanding of the correct storage conditions for the molecules in order to store them to be used for a long time.
2. Blotting Techniques
Blotting is the most commonly used laboratory technique that is extensively used in the area of cell and molecular biology. Blotting is an extremely effective multi-step procedure used in the analysis of biomolecules that are diverse, such as DNA, mRNA and proteins during various phases of gene expression or to identify particular proteins in a complex mix of proteins following extraction from cells. The applications are endless. Blotting techniques fall into the following kinds:
- Southern Blotting – widely used in gene mapping and discovery of genes, in the analysis of genetic patterns , and in Transgenic, Forensic & evolutionary research.
- Northern Blotting – gene expression analysis at the mRNA level as well as used in the analysis of RNA degradation, the splicing of RNA and determining its half-life.
- Western Blotting – identification of specific antibodies in complex, confirmatory test of HIV, Hepatitis B infection and Mad Cow Disease.
Other blotting techniques for alternative purposes are also available depending on the research requirements. With its broad range of uses, it is a essential lab skill for anyone who wants to make a profession in research. The mere knowledge of theory is not enough the hands-on training experiences with all types of blotting methods will let you carry out experiments and research easily in the course of your Ph.D. and at Work.
3. Gel Electrophoresis
Lab Techniques You Must Know For any analysis of molecular nature like analyzing DNA proteins and RNA must be processed using gel electrophoresis. Therefore, it is essential to master the electrophoretic equipment and to correctly cast the gel. The quality of the gel will give a clear and precise results. Top 10 lab techniques that every researcher should be aware of when making gel for casting, the most vital aspect is the precise proportion of the ingredient (agarose%/PA) because it will impact the size of the pore, that in turn determines the outcome of the experiment. Although agarose gel is used to analyze the structure of DNA PAGE (polyacrylamide gel electrophoresis) and SDS (Sodium Dodecyl Sulphate)-PAGE are used for protein analysis. The electrophoresis of agarose gel is the most efficient method to analyze DNA that is based on the size. Gel electrophoresis is extensively used in molecular biology labs and biochemistry in fields like conservation biology, forensic science and medical.
A few of the most significant uses of this technique are as follows:
- The separation of DNA fragments to be used for DNA fingerprinting to look into crime scene
- To examine the results of the polymerase chain reaction
- To study the genes that are that are associated with a specific illness
- In DNA profiling to aid taxonomy research to identify the different species
- Testing paternity by using DNA fingerprints
- The study of the protein structure and function
- The analysis of resistance to antibiotics
- Techniques for blotting to analyze of macromolecules.
- The study of evolutionary connections, we can study genetic similarities between species or populations
While this method of lab work is taught at the graduation level to all life science students, it is restricted. It is essential to be familiar with every aspect of Gel Electrophoresis procedure and the entire process it uses, its limitations, accuracy and application. The technique once you master it will stay with you until you’re in the field of research.
4. Bioinformatics Tools
In addition to being effective in wet-lab methods, researchers working in the area of genomics and proteomics requires a complete understanding of soft lab techniques also. To compare the homology of sequences – BLAST or to identify the structure of proteins – Rasmol or to create primersNEB cutters, and other tools must be mastered. Because the vast majority of proteomic and genomic research needs up-to-date information, scientists should be in routine practice of accessing databases of biotechnology and molecular research to find any information that has been updated. Bioinformatics in general is a broad field with numerous methods such as Molecular docking Molecular Modelling and Programming Languages such as – Python, C, C++, SQL, and the list is inexhaustible.
5. Flow Cytometry
This is a beneficial technique to sort cells i.e the process of separating different kinds of cells out from a heterogeneous mix of cells. This instrument is advanced enough that it is able to be used to detect various physical characteristics in a individual cell. The basis of the method is based on characteristics of light scattering in the particular cell that is identified by either monoclonal or dye-based antibodies on the cell’s surface or specific intracellular molecules.FACS permits the analysis of various cell populations within a short period of period of time. This device is crucial in cell biology as well as animal or plant tissue culture labs. So, anyone who is interested in a career in research in these areas should be able to gain hands-on experience with this method as soon as possible.
6. Chromatography Technique
Chromatography is founded on the idea of exploiting the partition effect which distributes the molecules across different phases, thereby breaking the complex mixture into components. In the majority of methods of chromatography, a stationary phase and a mobile phase is used to separate the molecules. Therefore, in the sense that chromatography is a technique used to distinguish biological molecules on the basis of their division between a stationary phase and a mobile and stationary phase.
- Thin layer Chromatography is used to isolate amino acids in proteins etc.Top 10 Laboratory Techniques Every Researcher Needs to be aware of
- Ion exchange chromatography is a widely used method of chromatography to separate the charged molecules like polypeptides and proteins nucleic acid, polypeptides, other molecules. The underlying principle behind ion exchange and chromatography is the exchange in two directions for the (ions) found in the solution with the ones electrostatically attached to the support medium.
- Affinity chromatography methods are founded on the separation of proteins since they are bound to specific receptors. is the particular interaction of the proteins with its receptors or ligands. Certain receptors or ligands, like for example, antigenic epitopes as well as those active enzyme sites show an extremely high degree of specificity to the protein in question, and affinity chromatography makes use from these interactions the purification of proteins in just one process.
- Size exclusion chromatography. Proteins are separated based on the size. As protein molecules pass through the gel, the smaller their size, the less they are stuck in the gel or sieve, and consequently that they will elute later. Larger molecules elute more quickly and are able to move faster, avoiding any tiny pores in the gel.
- High-Pressure Liquid Chromatography(HPLC): For HPLC the silica beads used in place of the polysaccharide beads. Due to the high pressure and the smaller sizes of silica beads this results in a more theoretical plates, resulting in greater resolution power to distinguish intricate biological samples. The reversed phase chromatography (RP-HPLC) makes use of resins that have tiny hydrophobic groups that are attached.
B.sc or B.tech or M.sc or M.tech Biotech or Life science applicants must be familiar with the protocol for a flow cytometry technique in your classes and, of course, gain a hands-on experience as well. You can join a Lab as an internship, or take an brief course or an internship on this subject. It’s not just an added benefit to your resume, but it will help you in your career in research or a future higher education.
7. Spectroscopic Techniques
UV-visible and mass spectroscopes can be generally used to conduct qualitative and quantitative research on biomolecules, like proteins. Nucleic acids etc. Ultraviolet is used in molecular biology as well as in microbial research because it is feasible to determine the quality, quantity and purity of nucleic acids and proteins and to determine the growth curve of microbial organisms. In the same way, mass spectroscopy is used for the measurement of molecular weights of proteins. NMR is used to refine structure and analysis of the structure of proteins to study the mechanisms behind secondary structure and function of a protein via folds, the conformational dynamic simulation of proteins and DNA as well as other. The chromatography techniques and their variants are listed below.
- UV-visible Quantitative and qualitative assay of nucleic acids, proteins and other substances.
- Circular Dichroism Secondary, Tertiary structures of proteins
- Atomic Absorption Spectrophotometer – Detection of metal Ions in biological samples
- The NMR structure of organic large molecules
- Ir Analyzing functional group in compounds
8. Reaction of Polymerase Chains
It is an extremely crucial methods in molecular biology. It is not just a groundbreaking technology, but is also extremely economical – by thousands of duplicates of DNA could be made using just a single DNA strand for a base. The Polymerase Chain Reaction method is extensively used in the medical and biochemical labs – is a essential component of nearly all kinds of tests. Some of its uses are as follows:
- DNA sequencing
- Genetic mutations are detected
- Cloning and expression profiling
- Tissue typing during organ transplantation procedures
- Genetic fingerprinting
- research on miRNA
- Testing for parental consent – in criminal cases
- Treatments for cancer that are formulated
- Early detection of all kinds of cancer and HIV
- DNA sequencing, cloning & gene expression
The applications listed here are only a small portion of the other. If you have a desire to make a impact in the world of research using this method, it can be a great help to you. The most advanced PCR devices are being developed by various manufacturers worldwide that you should know about these machines as well. Keep up-to-date your knowledge of the latest developments and developments in the field of the PCR machine, technique and use by reading research papers and trends in research.
9. Cell Culture Techniques
Lab Techniques You Must Know Culturing of bacterial as well as animal and plant cells is essential for microbial, cellular as well as molecular biological research. Microbial cultivation is one of the most effective methods to investigate the physiology, biochemistry and the genes of microbes, along with the screening for antibiotics. Additionally, microbes can be studied for the production of essential metabolites that could be used for production of the item on a vast scale. Microbes specialize in the breakdown of certain chemicals. Plant and animal cell culture is vital to study cellular metabolism, and is the most efficient method to study the effects on chemicals and drugs.
Cell culture is used in the study of mutagenesis as well as carcinogenesis through the use of powerful mutagens. Cells are also screened to discover new drugs and for development. Understanding this technique is an most important prerequisites for cell biology labs. Maintaining a clean environment using appropriate medium, incubation and subculturing of cells at the appropriate time is essential for the success of cell culture. Techniques for cell culture include the expertise of:
- Design and equipment to be used in Cell Culture Laboratory Cell Culture Laboratory
- Cell Culture is safe, but there are safety aspects to it.
- Sources of cell phone lines
- Cell Types and Characteristics of Culture
- Cryopreservation of cells and storage
- Cell Culture Protocols
- Identification of cell lines
10. Microscopic techniques
Being able to see clearly and in depth is crucial in the science field as in the field of science, for every declaration you make, it is essential that it is essential to have solid evidence from your experiments and the results that support your claims scientifically. Microscopes is used extensively in a vast array of subjects in science, specifically to comprehend the fundamentals of life and carry out vital research. With the continuous development in the microbiology area we are able to observe all kinds of cells, from plant and animal cells, to the structure of macromolecules within the cell.
Remember the first time in the laboratory of the college where you could see a stunning Top 10 Laboratory Techniques Every Researcher Should know about a stained plant cell in the microscope compound, and the smile it put to your face! It’s always thrilling and exciting every time to observe life on a micro-level It’s not it? It’s like a fluorescence microscope which is frequently used in the area of biomedical research. It assists in deducing details about the function of cells precisely through the emission of light by specific molecules labeled by a fluorescent dye. A particular field in science can’t make sense without a microscope, for instance the histology field. Histology is a extremely fascinating field that studies tissues at the microscopic scale. This field assists us to learn about functional morphology as as the dynamic of tissues. This is why this area is growing in popularity in its use in the area of cancer as well as similar illnesses.
The most modern technology, that is being used to build microscopes, the most intriguing ones are the ones that operate in real-time which provide live video footage of interactions between organisms and intracellular dynamics. Real-time microscopes also record these exciting events. Similar to in IISER, Pune Dr. Milind Watve had demonstrated how predation takes place in microbes by making use of a real-time microscope that also assisted him to comprehend the many new aspects that are arising around this concept. In looking back in the past important and complex scientific developments have been discovered using a microscope. To mention a few, here are some of the most interesting details of these events that were discovered using a microscope:
- Krebs Cycle written by Hans Krebs (1937)
- Cell division is a result of Walther Flemming (1879)
- Neurotransmission (between the end of the 19th century to the 20th century)
- Photosynthesis and cellular respiration discovered by Jan Ingenhousz in the 1770s
“An the eye of Science” a precise description is provided to the term microscope. Microscopes were, is an everlasting primary instrument of science, and therefore each researcher should be aware of having the most up-to-date understanding in that vast subject in order to improve and be able to comprehend science on a a tiny level.
11. Decolorization
The majority of organic compounds are in colorless. Highly conjugated substances (eg polymers with polarity) take up the light in the visible portion of the spectrum, and consequently appear “colored”. In the event that these extremely polar massive molecules are impurities they can be eliminated through the using finely granulated activated clay (Norit). Polar compounds (eg impurities that are polar) absorb to charcoal that is non-soluble in the solvent. They can then be removed from the solution. However there are some compounds that may also adsorb when there is enough charcoal . The technique is to select the proper amount. In most cases, a very small amount of charcoal is sufficient (there is a lot of surface area to the particles). It is added in tiny amounts into the warm (but still not the boiling) solution until a sufficient decolorization has taken place. Attention: trapped the air in the Norit could cause rapid frothing when it comes into contact with the solution that is hot. The Norit is able to be removed from the hot solution with filters made of fine paper, or a filters aid (Celite) that is laid over the filter paper. To accomplish this, create a Slurry of Celite in the solvent of your choice. Make sure the filter paper is wet, then apply suction to ensure it sticks. Then slowly pour the Celite slurry on the filter paper, making sure it covers it in a uniform manner. After that, apply the suction. The solvent can be removed from the flask in which you filter it or switch flasks for filter prior to filtration of the Norit-containing solution.
12. Crystallization
It is used to cleanse a solid. This process requires a appropriate solvent. A solvent that is suitable is one that is able to dissolve into the substance (solute) in the event that it is hot, but does not do so when it is cold. The best solvents have a significant distinction in solubility across a acceptable range of temperatures. (eg Water is a crystallization solvent that ranges from 100oC and 0oC. Hydrocarbon solvents like hexanes and petroleum ether possess a differing T range because they can be chilled to below 0 degrees, yet they will boil below 100oC.).
13. Centrifugation
The process of centrifugation is a mechanical procedure that makes use of centrifugal force to remove components from a solution based on their shape, size, the density of them, their medium viscosity, and the speed of the rotor. The more dense components of the mixture are displaced away from the direction of the centrifuge’s axis, and the lighter components of the mix move toward the center of the axis. Biologists and chemists can increase the gravitational force that is effective in the test tube to ensure that precipitate (pellet) can travel swiftly and completely towards at the base of the tube. The remaining liquid over the precipitate is known as a supernatant , also known as a supernate.
There is a connection between the density and size of a particle and the speed that it is separated from a heterogeneous mix when the sole force at work is gravity. The bigger the size and the higher the particle’s density and the quicker they are separated from the mix. When you apply a more powerful gravitational force on the mixture similar to what a centrifuge it, the separation of particles is speeded up. This is optimal in laboratory and industrial environments because particles that naturally break apart over a prolonged period of time are separated in shorter amounts of time.
The speed of centrifugation is determined by the angular speed, which is usually expressed in RPM, or revolutions per minute (RPM) or acceleration in terms of grams. The conversion rate between RPM and G is determined by the radius of the rotor of the centrifuge. The velocity of the particles’ settling in this process is a result of their size and shape, their centrifugal acceleration as well as the volume fraction of solids as well as the difference in density between the liquid and the particle as well as the degree of viscosity. The most commonly used use is that of separating solids from suspensions with high concentrations, and is used in treating sewage sludges to aid in dewatering , where less uniform sediment is created.
The method of centrifugation is used in a broad range of applications in both the labor and industrial sectors and not just is this procedure used to isolate two miscible substances as well as to study macromolecules’ hydrodynamic properties. This is an significant and widely used methods of research in biochemistry as well as molecular and cell biology. In the food and chemical industries, special centrifuges may handle a constant stream of particles-laden liquid. Centrifugation is the most commonly used method used for enriching uranium using the small mass differences between the atoms of U-238 as well as U-235 in the uranium-hexafluoride gas.
If you are a candidate who is interested in becoming extremely successful in their research careers have a thorough understanding of Bioinformatics tools is a necessity. The field is an emerging inter-disciplinary research field that is becoming increasingly used in all kinds of research, be it cancer research or mapping the human genome. So, start learning about Bioinformatics tools and techniques at an early stage in your profession.
Although these are the standard Lab techniques that researchers must be aware of, there are many more methods in the vast bioscience field, such as NGS, CRISPR, etc and knowledge about them is also attainable in the future.
Alongside these instrumental and lab methods, some basic laboratory procedures must be acquired. These techniques are simple and include autoclaving sterilization or cleaning the equipment as well as common preparations of reagents the use to use a centrifuge, a laminar airflow and preparation of media plating and the culture of microbes. They also include serial diluting, optimizing different equipment and water baths, creating cotton plugs for the culture tubes, and so on. These methods are typically learned after an experience in the laboratory. However, before launching an academic career a scientist must be technologically competent in every aspect.
