Molecular techniques ppt

All cooking is a special blend of science and art, but some styles go above and beyond to amaze your eyes and tastebuds. Molecular gastronomy uses techniques from chemistry and physics to craft edible creations that seem out of this world. Transforming the textures of food into innovative eating experiences, molecular gastronomy offers something new for the palates of intrepid diners.

From deconstructed desserts to transparent pasta and faux caviars, molecular gastronomy converts expectations into revelations on the dinner table. Using lab-like equipment such as liquid nitrogen and evaporators, this unique cooking style presents regular food in brand new ways that will blow your mind.

Would you try moss vapor? How about olive oil caviar, foam peas or powdered duck fat? Get familiar with common molecular gastronomy terms, so you can order with confidence when you see them on the menu.

Food prepared this way is always cooked evenly, with both the inside and outside equally tender. This allows the water inside fruits, vegetables and other fruits to freeze without creating large crystals or damaging the cell membranes, thus preserving the texture of frozen foods which would otherwise be mushy when defrosted.

Faux Caviar: Using a process known as spherification, liquid food like olive oil, tea and fruit juice can be turned into tiny little balls that look like caviar see top image. The liquid is held in its shape by a thin gel membrane and enjoyed as a solid. Deconstructed: If you deconstruct a sand castle, you knock it down.

This same idea applies to deconstructed dishes, which feature separate building blocks instead of having everything combined. Deconstructed dishes allow the diner to construct a customized experience in his or her mouth. Edible Paper: Made with potato starch and soybeans, these tasty sheets of paper are often printed with edible fruit inks from a laser printer.

Powdered Food: Chefs use maltodextrin, a starch-like substance, to turn a high-fat liquid like olive oil into a powder. Chefs are now turning fruits, vegetables and cheese into foams using food stabilizers and thickening agents. Image: criminalintent.

Molecular Techniques - PowerPoint PPT Presentation

Premium is the ad-free experience reserved for paying members. Support Organic Authority by subscribing to Premium and view the site with no ads.Hematopoietic disorders are often driven by genetic mutations and epigenetic alterations. New advanced technologies including next-generation sequencing, ultra-deep PCR and whole-genome and exome sequencing were proved very efficient in detecting several mutations implicated in the pathogenesis of hematological diseases.

Emerging evidence indicates that genomic data can be useful in all aspects of clinical practice including diagnosis, prognosis and prediction of response to specific treatments, as well as in the development of novel targeted treatments for patients with hematological disorders.

Benign and malignant hematological disorders are heterogeneous in both biological and clinical aspects. The alterations of genomic profile associated with these diseases are complex and variable including mutations, translocations, karyotypic rearrangements and post-translational modifications. In some cases, several genetic changes are required, to induce the onset of disease. This evidence in association with the evolution of molecular techniques has led to a modification of the existing dogma focusing on a single gene or single pathway analysis 1.

The development of new methods in molecular biology has not only allowed the individualized molecular diagnosis of diseases but has also led to the discovery of genetic or targeted therapeutic schemes with cytotoxic, anti-metabolic or immunomodulatory properties. Hematological physiology and pathology, independent of whether it is aggressive or indolent, affect patients of all ages with numerous clinical presentations. The concept of this review is to analyze the molecular basis of hematological diseases, as well as to present some new molecular technology and how they can affect overall survival.

Using polymerase chain reaction PCRkaryotype analysis, fluorescence in situ hybridization FISH and next-generation sequencing NGS it is possible to design better risk stratification categories and determine minimal residual disease MRD.

Immune check points inhibitors, antibodies and chimeric antigen receptor CAR -T cells can guide most efficient therapeutic strategies. Normal cell life is highly dependent on gene expression and any qualitative or quantitative alterations on the cascade of genetic information as well as changes in the time frame of gene activation, lead to inappropriate protein production 2.

Such changes induce irregular survival abilities, inappropriate response to external signals, autonomous amplification and deregulation of apoptosis pathway, formation of autocrine loops and promotion of angiogenic pathways. However, despite those irregularities, the mutated cell has a selective advantage.

Oncogenes refer to mutated genes generated from proto-oncogenes coding for proteins that regulate proliferation and differentiation or enhance epigenetic modifications. They are usually growth factors or mitogens secreted by cells with autocrine or paracrine properties.

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Such an example presents the c-Sis oncogene. Oncogenes may have the role of cytoplasmic tyrosine kinases. All these proteins constitute significant targets for treatment. The identification of Bcl - Abl fusion gene, was the first chromosomal abnormality associated with a specified disease and with the arrival of the Abl tyrosine kinase inhibitors TKIs in the history of CML turn over 56.

On the other hand, BTK is a tyrosine kinase involved in the transmission of different intracellular signals that reflect B cell physiology. Ras influences major signaling pathways that lead to cellular growth and proliferation. Finally, an oncogene may be a transcription factor, such as the myc gene, which regulates the transcription of genes that induce cellular proliferation. The detection of those chromosomal rearrangements expressed concomitant influence therapeutic strategies and reflect patient survival Tumor-suppressor genes encode for proteins that participate in the cell cycle.

They can be receptors for different growth factors or may play the role of enzymes that control DNA repair. Loss of expression of those genes is associated with high risk of developing a malignancy. The first tumor suppressor gene was identified by studies on retinoblastoma RB. The function of Rb as a tumor suppressor gene was validated by studies investigating the loss of normal Rb allele. Isolation of the Rb gene, as a molecular clone indemonstrated that Rb is lost or mutated in RBs.

Gene transfer experiments clarify that introduction of a normal Rb gene into RB cells cancels their tumorigenicity, indicating the activity of Rb as a tumor suppressor 14 Nodal role, in myeloproliferative and lymphoproliferative disorders, is played by the mutations in p53 protein, a nuclear transcription factor with a pro-apoptotic function, able to interrupt the cell cycle in G1 in response to damaged DNA and required for apoptosis induced by a variety of stimuli.

The mutations of p53 result in loss of function and are restricted within the DNA-binding domain of p They determine gene transcription via different mechanisms.Elizabeth A. Bello, Debra A. Anesthesiology ;85 6 We have emailed you at with instructions on how to set up a new password. If you do not receive an email in the next 24 hours, or if you misplace your new password, please contact:.

molecular techniques ppt

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molecular techniques ppt

Bello, MD ; Debra A. Schwinn, MD. Submitted for publication April I, Accepted for publication August 13, Address reprint requests to Dr.

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Address electronic mail to: schwi mc. Article Information. Review Article. Anesthesiology 12Vol. You will receive an email whenever this article is corrected, updated, or cited in the literature.

You can manage this and all other alerts in My Account. You must be logged in to access this feature.Recent revolutionary progress in human genomics is reshaping our approach to therapy and diagnosis. Nucleic acid—based testing is becoming a crucial diagnostic tool not only in the setting of inherited genetic disease e. Molecular diagnostics provides the necessary underpinnings for any successful application of gene therapy or biologic response modifiers.

It offers a great tool for assessing disease prognosis and therapy response and detecting minimal residual disease. Steps involved in a genetic approach to the diagnosis and treatment of disease. Reprinted with playing an integral role in the application of permission from reference 1. The newly established molecular pathology laboratory at Baylor University Medical Center positions our institution to provide state-of-the-art molecular testing as an integrated consultative element of our advanced patient care.

This, the first of a 2-part article, provides a general review of some principles and applications of molecular diagnostic techniques such as polymerase chain reaction PCRfluorescent in situ hybridization FISHspectral karyotype imaging SKIand DNA chip technology.

As an inherent part of oncogenesis, genetic rearrangements provide a great target for many molecular diagnostic tests in oncology. Rearrangements juxtapose otherwise distant segments of our genomic DNA. By bringing nucleic acid sequences closer together, new fusion chimeric genes are formed through chromosomal translocations or deletions of intervening DNA sequences. PCR is the most frequently used molecular technique in a molecular pathology laboratory.

Each PCR cycle involves 3 basic steps: denaturing, annealing, and polymerization. During annealing, or hybridization, oligonucleotide primers bind to their complementary bases on the single-stranded DNA.

The process is repeated 30 to 40 times, each cycle doubling the amount of the targeted genetic material. At the end of the PCR procedure, millions of identical copies of the original specific DNA sequence have been generated. Since these copies are identical in electrical charge as well as molecular weight, they are expected to migrate simultaneously, forming a single band, when applied to an electrophoretic gel.

Each cycle involves 3 steps: denaturing, annealing, and polymerization. During denaturing, the 2 strands of the helix of the target genetic material are unwound and separated by heating.

Finally, during polymerization, the polymerase enzyme reads the template strand and matches it with the appropriate nucleotides, resulting in 2 new identical helixes. After 30 to 40 cycles, millions of identical copies of the original DNA sequence are generated. If oligonucleotide primers used during the PCR cycles are labeled with fluorescent dye, the PCR product can then be analyzed in a capillary electrophoresis instrument, which tracks the fluorescence of the identical PCR sequences as they migrate.

A distinct peak indicates a positive amplification. The RNA sequence is first converted to a double-stranded nucleic acid sequence cDNA by using a reverse transcriptase enzyme borrowed from a retrovirus. As the name indicates, this technique allows for the real-time quantitation of PCR product following each of the 40 amplification cycles. The computerized Q-PCR instrument measures after each cycle the amount of fluorescence emitted from a dye intercalated in the double-helix DNA product; the amount of fluorescence is proportional to the number of copies of the amplification target.

When a certain critical copy number is reached, the amount of fluorescence increases by an exponential amount. Q-PCR therefore offers a great rapid quantitative advantage.

It is, moreover, less prone to contamination since the entire process of amplification and quantitation of the original target DNA for each sample is done in a single sealed tube. Q-PCR is of great utility in the assessment of minimal residual disease following novel targeted therapy against specific molecular defects as well as bone marrow transplantation for myelogenous leukemia. Not only can the presence or absence of leukemic cells carrying the target translocation t15;17; inv 16; or BCR-ABL now be evaluated, but a series of blood samples or bone marrow aspirates after transplantation can be compared to determine whether the number of BCR-ABL —positive cells in these samples is stable or is increasing.Cite This Article.

molecular techniques ppt

As molecular techniques for identifying and detecting microorganisms in the clinical microbiology laboratory have become routine, questions about the cost of these techniques and their contribution to patient care need to be addressed. Molecular diagnosis is most appropriate for infectious agents that are difficult to detect, identify, or test for susceptibility in a timely fashion with conventional methods. The tools of molecular biology have proven readily adaptable for use in the clinical diagnostic laboratory and promise to be extremely useful in diagnosis, therapy, and epidemiologic investigations and infection control 12.

Although technical issues such as ease of performance, reproducibility, sensitivity, and specificity of molecular tests are important, cost and potential contribution to patient care are also of concern 3.

Molecular methods may be an improvement over conventional microbiologic testing in many ways. Currently, their most practical and useful application is in detecting and identifying infectious agents for which routine growth-based culture and microscopy methods may not be adequate 4 — 7. Nucleic acid-based tests used in diagnosing infectious diseases use standard methods for isolating nucleic acids from organisms and clinical material and restriction endonuclease enzymes, gel electrophoresis, and nucleic acid hybridization techniques to analyze DNA or RNA 6.

Because the target DNA or RNA may be present in very small amounts in clinical specimens, various signal amplification and target amplification techniques have been used to detect infectious agents in clinical diagnostic laboratories 56.

Although mainly a research tool, nucleic acid sequence analysis coupled with target amplification is clinically useful and helps detect and identify previously uncultivatable organisms and characterize antimicrobial resistance gene mutations, thus aiding both diagnosis and treatment of infectious diseases 589. Automation and high-density oligonucleotide probe arrays DNA chips also hold great promise for characterizing microbial pathogens 6. Although most clinicians and microbiologists enthusiastically welcome the new molecular tests for diagnosing infectious disease, the high cost of these tests is of concern 3.

Despite the probability that improved patient outcome and reduced cost of antimicrobial agents and length of hospital stay will outweigh the increased laboratory costs incurred through the use of molecular testing, such savings are difficult to document 310 Much of the justification for expenditures on molecular testing is speculative 11 ; however, the cost of equipment, reagents, and trained personnel is real and substantial, and reimbursement issues are problematic 3 Given these concerns, a facility's need for molecular diagnostic testing for infectious diseases should be examined critically by the affected clinical and laboratory services.

In many instances, careful overseeing of test ordering and prudent use of a reference laboratory may be the most viable options.

Commercial kits for the molecular detection and identification of infectious pathogens have provided a degree of standardization and ease of use that has facilitated the introduction of molecular diagnostics into the clinical microbiology laboratory Table 1.

The use of nucleic acid probes for identifying cultured organisms and for direct detection of organisms in clinical material was the first exposure that most laboratories had to commercially available molecular tests. Although these probe tests are still widely used, amplification-based methods are increasingly employed for diagnosis, identification and quantitation of pathogens, and characterization of antimicrobial-drug resistance genes.

Commercial amplification kits are available for some pathogens Table 1but some clinically important pathogens require investigator-designed or "home-brew" methods Table 2. In addition, molecular strain typing, or genotyping, has proven useful in guiding therapeutic decisions for certain viral pathogens and for epidemiologic investigation and infection control 2A molecular marker is a DNA sequence in the genome which can be located and identified. As a result of genetic alterations mutations, insertions, deletionsthe base composition at a particular location of the genome may be different in different plants.

These differences, collectively called as polymorphisms can be mapped and identified. Plant breeders always prefer to detect the gene as the molecular marker, although this is not always possible.

The alternative is to have markers which are closely associated with genes and inherited together. The molecular markers are highly reliable and advantageous in plant breeding programmes:.

Let us assume that there are two plants of the same species—one with disease sensitivity and the other with disease resistance. If there is DNA marker that can identify these two alleles, then the genome can be extracted, digested by restriction enzymes, and separated by gel electrophoresis. The DNA fragments can be detected by their separation. For instance, the disease resistant plant may have a shorter DNA fragment while the disease — sensitive plant may have a longer DNA fragment Fig.

Marker-based DNA hybridization is widely used. The major limitation of this approach is that it requires large quantities of DNA and the use of radioactivity labeled probes. RFLP is mainly based on the altered restriction enzyme sites, as a result of mutations and re-combinations of genomic DNA. The procedure basically involves the isolation of genomic DNA, its digestion by restriction enzymes, separation by electrophoresis, and finally hybridization by incubating with cloned and labeled probes Fig.

Based on the presence of restriction sites, DNA fragments of different lengths can be generated by using different restriction enzymes. In the Fig. In plant A, a mutations has occurred leading to the loss of restriction site that can be digested by EcoRI. This results in a polymorphic pattern of separation. Locus non-specific markers e.

Locus specific markers e. Single short oligonucleotide primers usually a base primer can be arbitrarily selected and used for the amplification DNA segments of the genome which may be in distributed throughout the genome. The amplified products are separated on electrophoresis and identified. Based on the nucleotide alterations in the genome, the polymorphisms of amplified DNA sequences differ which can be identified as bends on gel electrophoresis.

Genomic DNA from two different plants often results in different amplification patterns i. This is based on the fact that a particular fragment of DNA may be generated from one individual, and not from others.

This represents polymorphism and can be used as a molecular marker of a particular species. Thus, this technique combines the usefulness of restriction digestion and PCR. The DNA of the genome is extracted. It is subjected to restriction digestion by two enzymes a rare cutter e.

Msel; a frequent cutter e. The cut ends on both sides are then ligated to known sequences of oligonucleotides Fig. PCR is now performed for the pre-selection of a fragment of DNA which has a single specific nucleotide. By this approach of pre-selective amplification, the pool of fragments can be reduced from the original mixture. In the second round of amplification by PCR, three nucleotide sequences are amplified. Autoradiography can be performed for the detection of DNA fragments.

Use of radiolabeled primers and fluorescently labeled fragments quickens AFLP. AFLP analysis is tedious and requires the involvement of skilled technical personnel. Hence some people are not in favour of this technique.

In recent years, commercial kits are made available for AFLP analysis.After you enable Flash, refresh this page and the presentation should play. Get the plugin now. Toggle navigation. Help Preferences Sign up Log in. To view this presentation, you'll need to allow Flash. Click to allow Flash After you enable Flash, refresh this page and the presentation should play.

View by Category Toggle navigation. Products Sold on our sister site CrystalGraphics. Title: Molecular Techniques. Tags: molecular techniques. Latest Highest Rated. Polymerase is activated. Primers dock or bookend target sequence, and polymerase docks to primer end. Light energy given off by the electrons movement is detected and measured, and the totality of fluorescence over cycles corresponds to the presence of the target nucleic acids. Photo detector Excitation?

Interfere with gene expression Bind with RNA - cause degradation block translation What else do they block???!!! Translocation e. Whether your application is business, how-to, education, medicine, school, church, sales, marketing, online training or just for fun, PowerShow. And, best of all, most of its cool features are free and easy to use. You can use PowerShow. Or use it to find and download high-quality how-to PowerPoint ppt presentations with illustrated or animated slides that will teach you how to do something new, also for free.

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molecular techniques ppt

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