This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
Genetic manipulation of plant and animal cells have to be confirmed for further application. One such confirmatory method is the use of stains/dyes which produces fluorescence when the recombination is successful.
Genomic library and shotgun sequencing. It includes the topics about genomic library,construction method, its uses and applications, shotgun sequencing, difference between random and whole genome sequencing, its advantages and disadvantages etc.
CLONING METHODOLOGIES:(GUYS LEARN CLONING IN EASIER WAY)
■PRINICIPLES AND STEPS INVOLVED IN CLONING
■METHODS INVOLVED IN cDNA OR GENOMIC CLONING
1.Isolation of mRNA
2.Synthesis of first strand of cDNA
3.Synthesis of second strand of cDNA
4.Cloning of cDNA
5.Introduction into Host Cell
6.Clone Selection
■OTHER TECHNIQUES INVOLVED IN CLONING OR FOREIGN GENE TRANSFER.
■EXPRESSION CLONING AND PROTEIN-PROTEIN INTERACTIONS
■cDNA or GENOMIC DNA LIBRARY CONSTRUCTION
■SIMILARITIES BETWEEN cDNA AND GENOMIC DNA LIBRARY
■ADVANTAGES AND DISADVANTAGES OF cDNA AND GENOMIC LIBRARIES
■REFERENCES.
Make my ppt useful in research and it also helpful for student's for Notes.
Genetic manipulation of plant and animal cells have to be confirmed for further application. One such confirmatory method is the use of stains/dyes which produces fluorescence when the recombination is successful.
Genomic library and shotgun sequencing. It includes the topics about genomic library,construction method, its uses and applications, shotgun sequencing, difference between random and whole genome sequencing, its advantages and disadvantages etc.
CLONING METHODOLOGIES:(GUYS LEARN CLONING IN EASIER WAY)
■PRINICIPLES AND STEPS INVOLVED IN CLONING
■METHODS INVOLVED IN cDNA OR GENOMIC CLONING
1.Isolation of mRNA
2.Synthesis of first strand of cDNA
3.Synthesis of second strand of cDNA
4.Cloning of cDNA
5.Introduction into Host Cell
6.Clone Selection
■OTHER TECHNIQUES INVOLVED IN CLONING OR FOREIGN GENE TRANSFER.
■EXPRESSION CLONING AND PROTEIN-PROTEIN INTERACTIONS
■cDNA or GENOMIC DNA LIBRARY CONSTRUCTION
■SIMILARITIES BETWEEN cDNA AND GENOMIC DNA LIBRARY
■ADVANTAGES AND DISADVANTAGES OF cDNA AND GENOMIC LIBRARIES
■REFERENCES.
Make my ppt useful in research and it also helpful for student's for Notes.
Class9 DNA technology in secondary schoolssusera700ad
Biotechnology is the use of an organism, or a component of an organism or other biological system, to make a product or process.
Many forms of modern biotechnology rely on DNA technology.
DNA technology is the sequencing, analysis, and cutting-and-pasting of DNA.
Common forms of DNA technology include DNA sequencing, polymerase chain reaction, DNA cloning, and gel electrophoresis.
Biotechnology inventions can raise new practical concerns and ethical questions that must be addressed with informed input from all of society.
DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine.
GENE CLONING,ITS HISTORY, NEW ADVENT IN GENE CLONING, PCR IMPORTANCE ,APPLICATION OF GENE CLONING,STEPS OF GENE CLONING,Antisense technology,Gene cloning in agriculture,Somatic cell therapy,Role of gene cloning in identification of genes responsible for human diseases,Synthesis of other recombinant human proteins and recombinant vaccines
Gene cloning in medicine,Recombinant protein from yeast,Problems with the production of recombinant protein in E.coli ,Expression of foreign genes in E.coli,Production of recombinant protein ,PCR can also be used to purify a gene,Obtaining a pure sample of a gene by cloning,Why gene cloning and PCR are so important,The advent of gene cloning and the polymerase
chain reaction.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
2. There are three main methods for obtaining genes
1. Synthesising the gene using an automated gene machine
2. Gene Cloning
3. Using the PCR
3. Gene Machine
•Recently, fully automated commercial instrument called automated polynucleotide
synthesizer or gene machine is available in market which synthesizes predetermined
polynucleotide sequence.
• Therefore, the genes can be synthesized rapidly and in high amount. For example, a
gene for tRNA can be synthesized within a few days through gene machine.
• It automatically synthesizes the short segments of single stranded DNA under the
control of microprocessor.
•The working principle of a gene machine includes (i) development of insoluble silica
based support in the form of beads which provides support for solid phase synthesis
of DNA chain, and (ii) development of stable deoxyribonucleoside phosphoramidites
as synthons which are stable to oxidation and hydrolysis, and ideal for DNA synthesis.
4. •Four separate reservoirs
containing nucleotides (A,T,C
and G) are connected with a
tube to a cylinder (synthesizer
column) packed with small
silica beads.
• These beads provide support
for assembly of DNA
molecules. Reservoirs for
reagent and solvent are also
attached.
•The whole procedure of
adding or removing the
chemicals from the reagent
reservoir in time is controlled
by microcomputer control
system i.e. microprocessor.
5. •If one desires to synthesize a short polynucleotide with a sequence of nucleotides
T,G,C, the cylinder is first filled with beads with a single 'T' attached.
• Thereafter, it is flooded with 'G' from the reservoir. The right hand side of each G
is blocked by using chemicals from the reservoir so that its attachment with any
other Gs can be prevented.
•The remaining Gs which could not join with Ts are flushed from the cylinder. The
other chemicals are passed from the reagent and solvent reservoirs so that these
can remove the blocks from G which is attached with the T.
•In the same way this cycle is repeated by flooding with C from reservoir into the
cylinder. Finally the sequence T.G.C is synthesized on the silica beads which is
removed chemically later on.
6. •The desired sequence is entered on a key board and the microprocessor
automatically opens the valve of nucleotide reservoir, and chemical and solvent
reservoir.
• In the gene machine the nucleotides are added into a polynucleotide chain at the
rate of two nucleotides per hour.
• By feeding the instructions of human insulin gene in gene machine, human insulin
has been synthesized.
7. Gene Cloning
•Gene cloning is a common practice in molecular biology labs that is used by
researchers to create copies of a particular gene for downstream
applications, such as sequencing, mutagenesis, genotyping or heterologous
expression of a protein.
• The traditional technique for gene cloning involves the transfer of a DNA
fragment of interest from one organism to a self-replicating genetic element,
such as a bacterial plasmid.
•This technique is commonly used today for isolating long or unstudied
genes and protein expression.
8. Steps in a gene cloning experiment are:
1 A fragment of DNA, containing the gene
to be cloned, is inserted into a circular
DNA molecule called a vector, to produce
a recombinant DNA molecule.
2 The vector transports the gene into a
host cell, which is usually a bacterium,
although other types of living cell can be
used.
3 Within the host cell the vector multiplies,
producing numerous identical copies, not
only of itself but also of the gene that it
carries.
4 When the host cell divides, copies of the
recombinant DNA molecule are passed to
the progeny and further vector replication
takes place.
5 After a large number of cell divisions, a
colony, or clone, of identical host cells is
produced. Each cell in the clone contains one
or more copies of the recombinant
DNA molecule; the gene carried by the
recombinant molecule is now said to be
cloned.
9. Polymerase Chain Reaction
•The polymerase chain reaction can also be used to obtain a pure sample of a gene.
•This is because the region of the starting DNA molecule that is copied during PCR is
the
segment whose boundaries are marked by the annealing positions of the two
oligonucleotide primers.
• If the primers anneal either side of the gene of interest, many copies of that gene
will be synthesized. The outcome is the same as with a gene cloning experiment,
although the problem of selection does not arise because the desired gene is
automatically “selected” as a result of the positions at which the primers anneal.
•An alternative to cloning, polymerase chain reaction(PCR), can be used to directly
amplify rare specific DNA sequences in a complex mixture when the ends of the
sequence are known.
10.
11. Genomic DNA is digested into large fragments using a restriction enzyme and then is
heat-denatured into single strands. Two synthetic oligonucleotides complementary to
the 3′ ends of the target DNA segment of interest are added in great excess to the
denatured DNA, and the temperature is lowered to 50 – 60 °C. The genomic DNA
remains denatured, because the complementary strands are at too low a concentration
to encounter each other during the period of incubation, but the specific
oligonucleotides, which are at a very high concentration, hybridize with their
complementary sequences in the genomic DNA. The hybridized oligonucleotides then
serve as primers for DNA chain synthesis, which begins upon addition of a supply of
deoxynucleotides and a temperature-resistant DNA polymerase such as that
from Thermus aquaticus (a bacterium that lives in hot springs). This enzyme, called Taq
polymerase, can extend the primers at temperatures up to 72 °C. When synthesis is
complete, the whole mixture is heated further (to 95 °C) to melt the newly formed DNA
duplexes. When the temperature is lowered again, another round of synthesis takes
place because excess primer is still present. Repeated cycles of synthesis (cooling) and
melting (heating) quickly amplify the sequence of interest. At each round, the number
of copies of the sequence between the primer sites is doubled; therefore, the desired
sequence increases exponentially.
12. Limitations of PCR:
•In order for the primers to anneal to the correct positions, either side of the gene
of interest, the sequences of these annealing sites must be known. It is easy to
synthesize a primer with a predetermined sequence, but if the sequences of the
annealing sites are unknown then the appropriate primers cannot be made. This
means that PCR cannot be used to isolate genes that have not been studied before—
that has to be done by cloning.
•There is a limit to the length of DNA sequence that can be copied by PCR.
Five kilobases (kb) can be copied fairly easily, and segments up to forty kb can be
dealt with by using specialized techniques, but this is shorter than the lengths of
many genes, especially those of humans and other vertebrates. Cloning must be
used if an intact version of a long gene is required.