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Experiome Organized a Guest Lecture

Experiome organized a Guest Lecture on August 27, 2022, entitled, Fundamentals of Recombinant DNA Technology. The lecture was delivered by Dr Chandra P Chaturvedi, Associate Professor, Department of Hematology, SGPGIMS, Lucknow. Team Experiome would like to sincerely thank Dr Chaturvedi for sharing his experiences and insights about the tools, techniques and applications of recombinant DNA technology

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Plant tissue culture generally refers to  in vitro culture of all plant parts under sterile conditions. Basic facilities are must in the laboratory where tissue culture technology is to be performed. This includes general cleaning and media preparation, sterilization, storage, sterile transfer, observation / data acquisition, and environmentally controlled incubator or culture room areas. The most important work area is the Cultural Transfer Office, where  core activities take place. Plant tissue culture should be cultured for a specified period of time under well-controlled temperature, humidity, airflow, and light quality. Plant culture techniques require a variety of organic and inorganic chemicals to prepare the  culture medium. The growing room is an equally important area where plant cultures are maintained under controlled environmental conditions for optimal growth. After in vitro culture of plants they are transferred to the pots. The potted plants are immediately transferred to the greenhouse or growth cabinets and are maintained under proper conditions of light, temperature, and humidity. Research and training organization

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Biotech Research Centre in Lucknow | Experiome Inc

We are a part of Vocal for Local, a government of India initiative. Join us for biotechnology training and research programs. Transforming Biotechnology Training landscape with Experiome Inc. Connect with us.. Biotech Research Centre in Lucknow | Biotech Training in Lucknow | Training Programs in Bioinformatics in Lucknow | Summer Training in Biotechnology and Bioinformatics in Lucknow

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Experiome organized a webinar, entitled “Perspectives on Genome Analysis and Editing: NGS and CRISPR-Cas, on 11th and 12th April, 2022, under the aegis of Vignan’s Foundation for Science, Technology & Research (Deemed University), Guntur, Andhra Pradesh.

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What is Genome Annotation?

The process of identifying the locations of genes and all of the coding sections in a

genome, as well as establishing what those genes perform, is known as genomeannotation. It's required because DNA sequencing generates sequences withuncertain functions.

Genome annotation

has progressed over the last three decadesfrom computational annotation of long protein-coding genes on single genomesand experimental annotation of short regulatory elements on a small number ofthem to population annotation of single nucleotides on thousands of individualgenomes.

Biotech Course

This improved precision and inclusivity of genome annotations (fromgenotypes to phenotypes) is resulting in more precise insights into the biology ofspecies, communities, and people.

Genome annotation

includes three major steps:

1. Identifying regions of the genome that do not encode proteins;

2. Identifying genome fundamentals, a process known as gene prediction; and

3. Identifying these elements' organic information

Annotation files include information about the genomic sequence. FASTA, GFF3,and GENBANK are few examples of file formats. There are various file formatsfor representing sequence, structure, and pathway information relating to genes andproteins, and internet databases provide the ability to select and download certainfiles.Genome annotation is of two types structural annotation that includes identificationof genomic elements while the assignment of function to structural elements isknown as functional annotation.

The genes or proteins that may be recruited by a certain genomic sequence can bepredicted using gene annotation algorithms. These new genes or proteins can befunctionally annotated by comparing their sequences to well-experimentallyconfirmed sequences in databases.Now that the genome sequences of over a thousand human persons (The 100,000Genomes Project, UK) and some model organisms are fully complete, genomeannotation remains a key hurdle for scientists exploring the human genome.Defining the biological "parts list" for the assembly and normal operation of anorganism is sometimes referred to as locating genes and other genetic regulatoryelements. Scientists are currently in the early stages of defining this parts list andfiguring out how all of the pieces "fit together.”

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Renato Dulbecco was the first to propose sequencing the human genome in 1984 to understand about cancer. The National Academy of Sciences in the United States first proposed HGP in 1988, followed by the National Institutes of Health and the Department of Energy. In 2003, the project was publicly established and declared. This project is still the largest collaborative biological effort in the world. Australia, Brazil, Canada, France, Germany, Japan, and the United Kingdom are among the countries participating in the initiative. It was a big problem to map out genes despite spending a large sum of money on it, starting at around $3 billion.

In addition, the project experts revealed that roughly 1.5 percent of DNA leads to the synthesis of proteins. This raises the question of what the other 98.5 percent of DNA, known as non-coding DNA, performs.

Over the last 20 years, scientists have had unprecedented access to the molecular basis of how a cell functions, not just in sickness but also in ordinary activities, thanks to the advancement of gene-splicing techniques. Scientists have mapped out the genetic molecules, or genes, that drive numerous life processes in ordinary microbes using these techniques. Researchers have begun to construct maps of human chromosomes, which contain many times the amount of genetic information as bacteria, thanks to continued advancements in biotechnology. These maps have led to the discovery of some significant genes, albeit in a primitive form. Short Term Biotechnology Training Centre in Lucknow.

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The requirement to test more proteins in fewer samples at the same time has prompted continued research towards increasing the sensitivity and speed of blotting techniques. Nonspecific interactions create false positives, which are eliminated by double blotting. Specific protein-protein interactions can be detected via Far-Western blotting. Proteins that interact with certain DNA sequences are identified by southwestern blotting. Multistrip blotting improves throughput while reducing variability between blots. New methods are being developed to lower the amount of protein required to produce a signal and to improve Western blotting's quantitative capabilities Summer Training in Biotechnology and Bioinformatics in Lucknow Experiome Inc.

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Experiome Inc-Biotech Research Centre in Lucknow | Biotech Training in Lucknow

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Next generation sequencing also known as high-throughput sequencing as it can sequence many DNA strands at the same time. This technology is used to study genetic variations associated with diseases by determining the sequence of DNA or RNA. Methods of Next generation sequencing are as follows:1.Massively Parallel Signature Sequencing2.Polony sequencing3.454 Pyrosequencing4.Ion Torrent5.Sequencing by Oligonucleotide Ligation Detection Life Science Training Centre in Lucknow | Biotech Research Centre in Lucknow | Biotech Training in Lucknow | Training Programs in Bioinformatics in Lucknow

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Next Generation Sequencing (NGS)

Next generation sequencing also known as high-throughput sequencing as it can sequence many DNA strands at the same time. This technology is used to study genetic variations associated with diseases by determining the sequence of DNA or RNA. Methods of Next generation sequencing are as follows:

1.Massively Parallel Signature Sequencing

This technology uses type II restriction endonuclease, a mixture of adapters with all possible overhangs which is annealed to the target sequence, leading in a single adapter with a perfect complimentary overhang being ligated. Since every adapter has its own label, the overhangs are recognised immediately after ligation. The number of oligonucleotide tags is 100 times greater than the number of template samples, ensuring that each template sample is conjugated to a unique tag. Signatures are generated by monitoring successful adapter ligations onto a surface of microbeads in a flow cell. This procedure uses millions of microbeads, and because each bead is connected to a single copy of the template, the sequencing reaction yields millions of signature sequences. Following that, cleavage using a type II restriction endonuclease exposes additional bases for identification in successive cycles. The fluorescent signals from the array of microbeads are recorded on a CCD camera, resulting in a digital representation of each microbead. Image-processing software then follows the positions of fluorescent signals from individual microbeads in the flow cell.

2.Polony sequencing

In it a universal linker or mate pair tags are used to circularise size-selected genomic segments. After that, universal sequences with complementary sections for sequencing or amplification primers are added to each fragment. Following emulsion PCR enrichment and immobilisation onto flow cells for monolayering, automated sequencing is conducted. During each sequencing cycle, four-color imaging is used to determine the sequence of each amplified bead at a specific location across several hundred raster points.

3.454 Pyrosequencing

Adapters are used to link fragmented template DNA to microbeads, followed by denaturation and a high dilution of the resulting library. Dilution is done to the point that each bead contains no more than a single DNA fragment. Individual DNA fragments are amplified using emulsion PCR after the DNA beads have been diluted. Emulsion PCR involves mixing beads containing DNA fragments, adapters, and PCR reagents with emulsion oil to create water droplets in an oil emulsion, resulting in droplets of water containing a single bead with a single DNA fragment attached. The PCR subsequently amplifies the DNA inside the water droplets in the oil solution, resulting in 106 double-stranded copies of the library segment in each bead. The amplified DNA-beads are then loaded into picotiter plates in such a way that each well contains only one bead and sequencing enzymes. Individual wells are packed with packing beads to assure the presence of a single bead in each well. The addition of one dNTP in each PCR cycle is then used to perform pyrrosequencing in a sequencing machine. When a proper nucleotide is incorporated, luciferase produces an oxyluciferin signal, which leads to the recognition of the individual nucleotides added to the nascent DNA. The sequence of the template fragment in each well is determined by the signals of each dNTP integrated, and the data of the resulting signals is pooled to give a sequence readout of all fragments.

4.Ion Torrent

Ion torrent chip has compartments and micro arrayed wells with solid state pH sensors. In it via emulsion PCR, sequence templates are generated on the beads resulting in oil-water emulsion spheres. An emulsion PCR results in the amplification of individual fragments to millions of identical copies that are linked to the beads, permits final detection of the signal. Each sphere includes one library molecule plus chemicals for the amplification. If an applied base is at that place in the template, a change in pH is noticed because individual dNTPs are applied in several cycles in a sequential order. When an identical stretch of bases is present, the pH changes by a factor of two, which is recognised and interpreted.

5.Sequencing by Oligonucleotide LigationDetection

It consists of fragmented DNA samples, end repair, and ligation of two different DNA adapters to the ends of the library fragments. The capacity of DNA ligase to recognise and integrate bases in a very particular manner is the basis of this sequencing. DNA fragments connected to beads are clonally amplified by emulsion PCR in sequencing by ligation. Following PCR, specific primers hybridise to the adaptor sequence of the amplified templates on the beads, resulting in a free 5′ phosphate group for ligation to the fluorescently labelled probes (known as interrogation probes) rather than a 3′ hydroxyl group as in traditional polymerase-mediated extension. The interrogation probe is 8 bp long, with the first two bases being specific and the next six bases being degenerate. At the end, a set of four fluorescently labelled interrogation probes compete for ligation to the sequencing primer, each of which has one of 16 potential 2-base combinations. When a probe is ligated, fluorescence is collected that corresponds to the probe ligated. The connected probe's "fluor" is eliminated in the second cycle, and a 5′ phosphate group is regenerated. Multiple ligation, detection, and cleavage cycles are carried out, with the number of cycles defining the final read length. The extension product is removed after a set of ligation cycles (typically seven), and the template is reset with a primer complementary to the n-1 location for another set of ligation cycles. Each time, a new primer with a different offset is used to continue the procedure.

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At Experiome, we give outstanding importance on excellent training in our thoroughly operational biotechnology laboratory.

The training modules are designed keeping in view the continuously evolving biotechnology education and research scenario of the country.

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Biotech Training in Lucknow | Experiome

Why Biotechnology trainingis important and relevant to the students of PG and UG?

Biotechnology consists of altering biological function by adding or removing genes from another organism. This course isintendedfor students with abackgroundin biology and nodding acquaintance in maths and chemistry till class 12. After graduation it is necessary to pass national level examinations to be part of academics. Apart of this biotechnology has a significant impact on a variety of industries, including biomedical, agriculture, health care, farming, food processing, the environment, chemicals, and textiles. Considering the situation, biotechnology has grown in popularity over time, and in the not-too-distant future, it will reach the skies. So, what really is the significance of training? It is critical for students to have hands-on training and understanding in the area. According to the current situation, freshers do not receive adequate hands-on training, resulting in job loss. To bridge the gap between academic knowledge and industrial training, biotechnology training is required. This is gained through training in well-equipped and knowledgeable labs. These trainings are beneficial because students receive understanding of how to operate industry tools, giving them the confidence to work in any lab or business. Aside from that, industrial training will improve students' style of thinking, build technical abilities, and presentation skills, as well as help them grasp science better and perform better in interviews.

A huge number of students have been drawn to the field of biotechnology, and each year a considerable number of students graduate with B.Sc./ M.Sc., B.Tech/ M.Tech degrees. For R&D, production, and services, the biotechnology business requires trained labour. The majority of students, however, are unable to find work since they lack the skills required by the biotech industry. In addition, the biotech industry has stated on multiple occasions that it lacks competent labour to meet its needs. There is a pressing demand for B.Sc., B.Tech., and M.Sc., M.Tech. candidates who want to specialize in the biotech industry to receive skills training. Due to the students' core knowledge, extra abilities can be delivered in a short period of six months through theoretical and hands-on exposure.

We at Experiome guide and direct students about the career and provide them with the latest hands-on training in the field of biotechnology, bioinformatics, biochemistry and other areas of life sciences.

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CRISPR-Cas use to meet the need of Vitamin A

Nutrition is essential for healthy growth and development. Macronutrients or micronutrients are the two types of nutrients. Requirements of macronutrients including fats, proteins, and carbohydrates are relatively higher to that of micronutrients including several vitamins and minerals. In comparison, if the amount of micronutrients in the diet is low, it has an impact on the child's physical and mental development.

Vitamin A is a lipid-soluble vitamin that is required for cell growth, metabolism, immunological function, eyesight, and reproduction.It is absorbed in the duodenum and stored in hepatic stellate cells, as well as in adipose tissue and the pancreas in small amounts. Vitamin A is found naturally in dark leafy greens, orange-colored vegetables, milk products, liver, and fish. Vitamin A intake should be around 200 micrograms per day for children aged 1 to5, 700 micrograms per day for women, and 900 micrograms per day for males.

Vitamin A deficiency is a source of concern for children in impoverished countries. According to UNICEF, one-third of children do not receive vitamin A supplementation, while the WHO estimates that 190 million pre-school children are vitamin A deficient. According to a survey conducted by the National Nutrition Monitoring Bureau (NNMB), around 62 percent of preschool children in India have Vitamin A deficiency, and 21.5 percent have serum retinol levels of less than 0.35 mol/l.

Considering the scenario, biotechnologists areattempting to enhance the Vitamin Acontent by using clustered regularly interspaced short palindromic repeat (CRISPR) technology. Banana is economically important fruit throughout the world. Genetically modified crops with the help of CRISPR-Cas are found to be promising. Recently, the carotene-rich Cavendish banana cultivar (cv.) Grand Naine was developed using a CRISPR/Cas9-based method (AAA genome). The lycopene epsilon-cyclase (LCYε) gene's fifth exon was targeted. Designed guide-RNA’s targeting specificity was also tested by its capacity to develop indels in the LCYε gene in cv. Rasthali (AAB genome) genome. In comparison to control (without genome editing) plants, metabolic analysis of the fruit pulp of the chosen edited lines revealed a 6-fold increase in beta-carotene concentration (24 microgram/g). Enhancing the quality and the content of the fruit via CRISPR could be a potential alternative for the population suffering from Vitamin A deficiency.

We, at Experiome, offer training in CRISPR-Cas vector designing. We provide bioinformatics support for CRISPR-Cas vector design for any gene of interest.

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CRISPR-Casuse to meet the need of Vitamin A

Nutrition is essential for healthy growth and development. Macronutrients or micronutrients are the two types of nutrients. Requirements of macronutrients including fats, proteins, and carbohydrates are relatively higher to that of micronutrients including several vitamins and minerals. In comparison, if the amount of micronutrients in the diet is low, it has an impact on the child's physical and mental development.

Vitamin A is a lipid-soluble vitamin that is required for cell growth, metabolism, immunological function, eyesight, and reproduction. It is absorbed in the duodenum and stored in hepatic stellate cells, as well as in adipose tissue and the pancreas in small amounts. Vitamin A is found naturally in dark leafy greens, orange-colored vegetables, milk products, liver, and fish. Vitamin A intake should be around 200 micrograms per day for children aged 1 to5, 700 micrograms per day for women, and 900 micrograms per day for males.

Vitamin A deficiency is a source of concern for children in impoverished countries. According to UNICEF, one-third of children do not receive vitamin A supplementation, while the WHO estimates that 190 million pre-school children are vitamin A deficient. According to a survey conducted by the National Nutrition Monitoring Bureau (NNMB), around 62 percent of preschool children in India have Vitamin A deficiency, and 21.5 percent have serum retinol levels of less than 0.35 mol/l.

Considering the scenario, biotechnologists areattempting to enhance the Vitamin Acontent by using clustered regularly interspaced short palindromic repeat (CRISPR) technology. Banana is economically important fruit throughout the world. Genetically modified crops with the help of CRISPR-Cas are found to be promising. Recently, the carotene-rich Cavendish banana cultivar (cv.) Grand Naine was developed using a CRISPR/Cas9-based method (AAA genome). The lycopene epsilon-cyclase (LCYε) gene's fifth exon was targeted. Designed guide-RNA’s targeting specificity was also tested by its capacity to develop indels in the LCYε gene in cv. Rasthali (AAB genome) genome. In comparison to control (without genome editing) plants, metabolic analysis of the fruit pulp of the chosen edited lines revealed a 6-fold increase in beta-carotene concentration (24 microgram/g). Enhancing the quality and the content of the fruit via CRISPR could be a potential alternative for the population suffering from Vitamin A deficiency.

We, at Experiome, offer training in CRISPR-Cas vector designing. We provide bioinformatics support for CRISPR-Cas vector design for any gene of interest.

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We work on a two-pronged philosophy of knowledge dissemination and knowledge creation, and our research solutions work diligently towards knowledge creation. Even our training modules are oriented towards research thought or ideas that can be published or work as preliminary data for further work.

We provide research services and support in the areas of cloning, in silico analysis, microarray and next generation sequencing (NGS) data analyses, development of NGS data analysis workflows and pipelines, etc. We work closely with our clients, and provide tailor-made solutions that seamlessly integrate with their vision.

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Summer Training in Biotechnology and Bioinformatics | Biotech Training-Experiome

Experiome

is organising a 3-day Hands on Skill Development Workshop in Biotechnology

Spirulina is a blue-green algae or cyanobacteria which is found in high-salt alkaline water, subtropical and tropical places such as America, Mexico, Asia, and Central Africa.  It’s asexual reproduction reduces the danger of gene escape into the food chain, as well as the accompanying food security and regulatory concerns. It contains a lot of macro and micronutrients, important amino acids, proteins, lipids, vitamins, minerals, and antioxidants. With these characteristics, photosynthetic cyanobacteria have been used in a variety of industries, including food, health, and pharmaceuticals. It is consumed mainly in central Africa and later NASA started using spirulina as a nutritional supplement for astronauts on space missions. It gained attention for therapeutic benefits like hypercholesterolemia, hyperglycerolemia, cardiovascular diseases, inflammatory diseases, allergies, cancer and viral infections.

Spirulina, as a plant-based biopharmaceutical, has the potential to overcome the constraints and limits of existing crop-and algal-based platforms.

Contact for more information: 9839534061, 7266991117

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Spirulina is a blue-green algae or cyanobacteria which is found in high-salt alkaline water, subtropical and tropical places such as America, Mexico, Asia, and Central Africa.  It’s asexual reproduction reduces the danger of gene escape into the food chain, as well as the accompanying food security and regulatory concerns. It contains a lot of macro and micronutrients, important amino acids, proteins, lipids, vitamins, minerals, and antioxidants. With these characteristics, photosynthetic cyanobacteria have been used in a variety of industries, including food, health, and pharmaceuticals. It is consumed mainly in central Africa and later NASA started using spirulina as a nutritional supplement for astronauts on space missions. It gained attention for therapeutic benefits like hypercholesterolemia, hyperglycerolemia, cardiovascular diseases, inflammatory diseases, allergies, cancer and viral infections.

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