Nucleic Acid Hybridization.pptx
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CONTENT -Hybridization Introduction -Southern Hybridization Blotting (Transfer Of DNA On A Membrane) -Application -DNA Fingerprinting -DNA Typing Introduction-Hybridization The capacity of denatured DNA to reanneal under appropriate ionic strength and temperature is known as Hybridization. In case of a mixture of DNA molecules, hybridization leads to formation of hybrid molecules. Often a DNA probe is used in hybridization for detecting or finding a particular DNA sequence from a mixture of DNA molecules. Hybridization may be done with DNA fragments separated by electrophoresis, dot blot: DNA spots on a membrane or colony blot: hybridization with bacterial colonies containing particular rDNA. Southern Hybridization A method of DNA hybridization established by Ed Southern in 1975. Genomic DNA from an organism is restriction digested and separated by electrophoresis. The electropherosed DNA is transferred on to a membrane. The membrane bound DNA is denatured using an alkali. A DNA probe is radiolabeled and added to the denatured DNA in presence of appropriate buffer for hybridization. After sufficient time for hybridization the membrane is exposed to X-ray film. The X-ray film shows signals of hybridization: where the DNA probe has complementary structure on the transferred DNA. The resulting X-ray film is known as an Autoradiogram and the process Autoradiography. DNA fingerprinting Using the method of DNA hybridization with an appropriate DNA probe to identify individuals as done with fingerprinting. The method was first done by Sir Alec Jeffreys in 1985. Any biological material such as a drop of blood, saliva, semen, and any body part such as bones, tissue, skull, teeth, hair with root etc found at the scene of crime is used as source DNA. DNA probe used for DNA fingerprinting may be a VNTR or minisatellite DNA. The probe is labeled and used to hybridize the source DNA. An autoradiogram is generated that show different band patterns for different individuals.
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Genetic Engineering and Genomics Notes - MH-CET 2015
Genetic Engineering and Genomics Notes - MH-CET 2015
Blotting techniques includes southren,northern,western and dot blotting
Blotting techniques includes southren,northern,western and dot blotting
The power of dna fingerprinting in forensic science
The power of dna fingerprinting in forensic science
southern blot, western blot,blotting technique ,blotppt.pptx
southern blot, western blot,blotting technique ,blotppt.pptx
More from Nusrat Sheikh
Random amplified polymorphic DNA-RAPD.pptx
Random Amplified Polymorphic DNA is a type of PCR in which the segments of DNA that are amplified are random.
Williams et al. (1990) developed this technique using very short primers to generate random fragments from template DNAs.
RAPD fragments can be separated and used as genetic markers or a kind of DNA fingerprinting.
Principle
The standard RAPD technology utilizes short synthetic oligonucleotides (10 bases long) of random sequences as primers to amplify nanogram amounts of total genomic DNA under low annealing temperatures by PCR. Amplification products are generally separated on agarose gels and stained with ethidium bromide.
At an appropriate annealing temperature during the thermal cycle, oligonucleotide primers of random sequence bind several priming sites on the complementary sequences in the template genomic DNA and produce discrete DNA products if these priming sites are within an amplifiable distance of each other.
The profile of amplified DNA primarily depends on nucleotide sequence homology between the template DNA and oligonucleotide primer at the end of each amplified product. Nucleotide variation between different sets of template DNAs will result in the presence or absence of bands because of changes in the priming sites.
Procedure
1. DNA is made single stranded by raising the temperature to 940C.
2. In second step, temperature is lowered to 40- 650C which results in annealing of the primer to their target sequences on the template DNA.
3. Temperature is chosen where the activity of the thermostable Taq DNA polymerase is optimal.
4. The polymerase now extends the 3’ ends of the DNA primer hybrids towards the outer primer binding site.
5. Repeating these three step cycles 40 to 50 times results in the exponential amplification of the target between the 5’ ends of the two primer binding sites.
6. Amplification products are separated by gel electrophoresis and visualized by ethidium bromide staining.
NORTHERN BLOTTING.pptx
This document describes the process of northern blotting. Northern blotting is used to detect specific RNA molecules and analyze gene expression. It involves separating RNA samples by size using gel electrophoresis, transferring them to a membrane, then using a labeled probe to detect the RNA of interest. The key steps are isolating RNA from tissue or cells, separating the RNA by size on a gel, blotting the RNA onto a membrane, hybridizing the membrane with a labeled probe, washing off excess probe, and visualizing where the probe bound to detect the RNA of interest. Northern blotting allows researchers to study gene expression patterns in different tissues, developmental stages, or disease states.
southern blotting.pptx
Blotting
A blot, in molecular biology and genetics, is a method of transferring proteins, DNA or RNA, onto a carrier.
The term "blotting" refers to the transfer of biological samples from a gel to a membrane and their subsequent detection on the surface of the membrane.
Types of blotting techniques
Southern Blotting
Northern Blotting
Western Blotting
A Southern blot is a method used
in molecular biology for detection of a specific DNA sequence in DNA samples.
Southern blotting combines transfer of electrophoresis -separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization.
The method is named after its inventor, the British biologist Edwin Mellor Southern.
- Methods in Southern blotting
- Advantages and disadvantages
Vernalization in Plants.pptx
Introduction:
Vernalization is the process whereby flowering is promoted by a cold treatment given to a fully hydrated seed or to a growing plant.
Dry seeds do not respond to the cold treatment.
Due to vernalization, the vegetative period of the plant is cut short resulting in an early flowering.
Also called yarovization.
Without the cold treatment, plants that require vernalization show delayed flowering or remain vegetative.
In many cases, these plants grow as rosettes with no elongation of the stem.
History:
Klippart,1857- first noticed the low-temperature requirement for flowering while working with winter wheat and spring wheat.
Lysenko,1938-used the term vernalization for a low-temperature promotion of flowering in plants.
Chourad ,1960- defined vernaliZation as “acquisition or acceleration of the ability to flower by a chilling treatment”.
Vernalization
For vernalization the seeds are allowed to germinate for some time and then are given cold treatment from 0 ̊C to 5 ̊C.
The period of cold treatment, varies from few days to many weeks.
After the cold treatment the seedlings are allowed to dry for some time and then sown.
Vernalization prepares the plant for flowering.
The cold stimulus usually perceived by the apical meristems. But in some species, all dividing cells of roots and leaves may be the potential sites of vernalization eg. Leennario biennis.
Vernalization induces the plant to produce a hormone called vernalin. It was discovered by Melcher(1936).
The vernalization stimulus can be transmitted from one plant to another through grafting.
The age of the plant is an important factor in determining the responsiveness of the plant to the cold stimulus and it differs in different species.
The suitable temperatures for vernalization ranges between 1 to 6 ̊c.
At higher temperature from 7 ̊c onwards response of the plant is decreased.
A temperature of about 12 to 14 ̊c is most ineffective in vernalizing the plant.
The vernalization is an aerobic process and requires metabolic energy.
In the absence of oxygen cold treatment becomes completely inefficient.
A sufficient amount of water is also essential.
Vernalization of dry seeds is not possible.
Factors affecting Vernalization:
Site of vernalization
Age of plants
Appropriate low temperature
Duration of exposure
Mechanism of vernalization:
Two theories..
1. Phasic development theory
2. Hormonal theories.
Epigenetic Changes in Gene Expression:
Vernalization May Involve Epigenetic Changes in Gene Expression.
Changes in gene expression that are stable even after the signal that induced the change (in this case cold) is removed are known as epigenetic regulation.
One model for how vernalization affects flowering is that there are stable changes in the pattern of gene expression in the meristem after cold treatment.
The involvement of epigenetic regulation in the vernalization process has been confirmed in the LDP Arabidopsis.
Photoperiodism
PHOTOPERIODISM IN PLANTS
• The concept of photoperiodism was given by W.W. Garner & H.A. Allard of the U.S Department of Agriculture, studied flowering in Maryland's mammoth variety of Tobacco plant in 1920.
• ‘PHOTOPERIODISM':-derived from 2 Greek words i.e.,'photos' (light) & periods (length of time).
• With a few exceptions, virtually all organisms (plant & animals) need exposure to light for a given number of hours per day for a variety of growth and reproductive functions. This day length is called PHOTOPERIOD & the phenomenon is called PHOTOPERIODISM.
• The flowering plant comprises 2 phases i.e, vegetative & reproductive.
• Under an appropriate photoperiod, plant switches from vegetative to reproductive phase, responding to the synthesis of flowering hormones & its subsequent translocation to buds. (Bartholomew,1977)
• The effect of daily duration of light hours(& dark periods) on the growth & development of plants, especially flowering is called photoperiodism.
CRITICAL DAY LENGTH
• Critical day length is the photoperiod required to induce flowering.
• It varies from species to species.
• e.g-Xanthium (SDP) requires a critical day length of 15.5hrs (15.5 light/8.5 dark).
• Critical photoperiod mustn't be exceeded in short-day plants & should always be exceeded in long-day plants.
• A single photoperiodic cycle that induces flowering-Inductive cycle & its effect is called Photoperiodic induction.
Depending upon the duration of the photoperiod, plants are classified into 3 categories:
1. Short Day Plants (SDP):
These plants require a relatively short daylight period (usually 8-10 hours) and a continuous dark period of about 14-16 hours for subsequent flowering.E.g.:-Strawberry, coffee, pineapple, etc.
o These plants are also known as long-night-plants.
2. Long day plants (LDP):
These plants require a longer daylight period (usually 14-16 hours) in a 24 hours cycle for subsequent flowering.
o Also called as short night plants.
E.g.:-Apple, passion fruit, etc..
3. A day-neutral plant (DNP):
This plants flower in all photoperiods ranging from 5 hrs to
24 hrs of continuous exposure.
e.g.:- Banana, guava, tomato, brinjal, etc…
-Dual Day Length Plants
Long Short Day Plants(LSDP):
These are short-day plants but must be exposed to long days during early periods of growth for subsequent flowering.
e.g.:- species of Bryophyllum, night jasmine, etc..
Short-Long Day Plants(SLDP):
These are long-day plants but must be exposed to short days during early periods of growth for subsequent flowering.
e.g.:- Wheat (Triticum), Rye (Secale), etc.
-MECHANISM OF PHOTOPERIODISM
-Florigen concept
-Phytochrome concept
-IMPORTANCE OF PHOTOPERIODISM
Biosynthesis of phospholipids
BIOSYNTHESIS OF PHOSPHOLIPIDS
Phospholipids:-
These are compounds containing, in addition to fatty acid and glycerol, phosphoric acid, nitrogenous bases, and another substituent. Polar compounds composed of alcohol attached by phosphodiester bridge to either diacylglycerol or sphingosine.
Amphipathic in nature has a hydrophilic head (phosphate +alcohol
eg., serine, ethanolamine, and choline) and a long, hydrophobic tail
(fatty acids or derivatives ).
- CLASSIFICATION OF PHOSPHOLIPIDS:-
- Glycerophospholipids
- Spingophospholipids or Sphingomyelin
- SYNTHESIS OF PHOSPHOLIPIDS
- FUNCTIONS OF PHOSPHOLIPIDS
- FUNCTIONS OF SPHINGOLIPIDS
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https://ashpc.eu/
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Presented at June 6-7 Texas Alliance of Groundwater Districts Business MeetingDescribing and Interpreting an Immersive Learning Case with the Immersion Cub...
Current descriptions of immersive learning cases are often difficult or impossible to compare. This is due to a myriad of different options on what details to include, which aspects are relevant, and on the descriptive approaches employed. Also, these aspects often combine very specific details with more general guidelines or indicate intents and rationales without clarifying their implementation. In this paper we provide a method to describe immersive learning cases that is structured to enable comparisons, yet flexible enough to allow researchers and practitioners to decide which aspects to include. This method leverages a taxonomy that classifies educational aspects at three levels (uses, practices, and strategies) and then utilizes two frameworks, the Immersive Learning Brain and the Immersion Cube, to enable a structured description and interpretation of immersive learning cases. The method is then demonstrated on a published immersive learning case on training for wind turbine maintenance using virtual reality. Applying the method results in a structured artifact, the Immersive Learning Case Sheet, that tags the case with its proximal uses, practices, and strategies, and refines the free text case description to ensure that matching details are included. This contribution is thus a case description method in support of future comparative research of immersive learning cases. We then discuss how the resulting description and interpretation can be leveraged to change immersion learning cases, by enriching them (considering low-effort changes or additions) or innovating (exploring more challenging avenues of transformation). The method holds significant promise to support better-grounded research in immersive learning.
Bob Reedy - Nitrate in Texas Groundwater.pdf
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The binding of cosmological structures by massless topological defects
Assuming spherical symmetry and weak field, it is shown that if one solves the Poisson equation or the Einstein field
equations sourced by a topological defect, i.e. a singularity of a very specific form, the result is a localized gravitational
field capable of driving flat rotation (i.e. Keplerian circular orbits at a constant speed for all radii) of test masses on a thin
spherical shell without any underlying mass. Moreover, a large-scale structure which exploits this solution by assembling
concentrically a number of such topological defects can establish a flat stellar or galactic rotation curve, and can also deflect
light in the same manner as an equipotential (isothermal) sphere. Thus, the need for dark matter or modified gravity theory is
mitigated, at least in part.
在线办理(salfor毕业证书)索尔福德大学毕业证毕业完成信一模一样
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四.办理各国各大学文凭(一对一专业服务,可全程监控跟踪进度)
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Immersive Learning That Works: Research Grounding and Paths Forward
We will metaverse into the essence of immersive learning, into its three dimensions and conceptual models. This approach encompasses elements from teaching methodologies to social involvement, through organizational concerns and technologies. Challenging the perception of learning as knowledge transfer, we introduce a 'Uses, Practices & Strategies' model operationalized by the 'Immersive Learning Brain' and ‘Immersion Cube’ frameworks. This approach offers a comprehensive guide through the intricacies of immersive educational experiences and spotlighting research frontiers, along the immersion dimensions of system, narrative, and agency. Our discourse extends to stakeholders beyond the academic sphere, addressing the interests of technologists, instructional designers, and policymakers. We span various contexts, from formal education to organizational transformation to the new horizon of an AI-pervasive society. This keynote aims to unite the iLRN community in a collaborative journey towards a future where immersive learning research and practice coalesce, paving the way for innovative educational research and practice landscapes.
ESR spectroscopy in liquid food and beverages.pptx
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...
The pygmy halfbeak Dermogenys colletei, is known for its viviparous nature, this presents an intriguing case of relatively low fecundity, raising questions about potential compensatory reproductive strategies employed by this species. Our study delves into the examination of fecundity and the Gonadosomatic Index (GSI) in the Pygmy Halfbeak, D. colletei (Meisner, 2001), an intriguing viviparous fish indigenous to Sarawak, Borneo. We hypothesize that the Pygmy halfbeak, D. colletei, may exhibit unique reproductive adaptations to offset its low fecundity, thus enhancing its survival and fitness. To address this, we conducted a comprehensive study utilizing 28 mature female specimens of D. colletei, carefully measuring fecundity and GSI to shed light on the reproductive adaptations of this species. Our findings reveal that D. colletei indeed exhibits low fecundity, with a mean of 16.76 ± 2.01, and a mean GSI of 12.83 ± 1.27, providing crucial insights into the reproductive mechanisms at play in this species. These results underscore the existence of unique reproductive strategies in D. colletei, enabling its adaptation and persistence in Borneo's diverse aquatic ecosystems, and call for further ecological research to elucidate these mechanisms. This study lends to a better understanding of viviparous fish in Borneo and contributes to the broader field of aquatic ecology, enhancing our knowledge of species adaptations to unique ecological challenges.
20240520 Planning a Circuit Simulator in JavaScript.pptx
Evaporation step counter work. I have done a physical experiment.
(Work in progress.)
Authoring a personal GPT for your research and practice: How we created the Q...
Thematic analysis in qualitative research is a time-consuming and systematic task, typically done using teams. Team members must ground their activities on common understandings of the major concepts underlying the thematic analysis, and define criteria for its development. However, conceptual misunderstandings, equivocations, and lack of adherence to criteria are challenges to the quality and speed of this process. Given the distributed and uncertain nature of this process, we wondered if the tasks in thematic analysis could be supported by readily available artificial intelligence chatbots. Our early efforts point to potential benefits: not just saving time in the coding process but better adherence to criteria and grounding, by increasing triangulation between humans and artificial intelligence. This tutorial will provide a description and demonstration of the process we followed, as two academic researchers, to develop a custom ChatGPT to assist with qualitative coding in the thematic data analysis process of immersive learning accounts in a survey of the academic literature: QUAL-E Immersive Learning Thematic Analysis Helper. In the hands-on time, participants will try out QUAL-E and develop their ideas for their own qualitative coding ChatGPT. Participants that have the paid ChatGPT Plus subscription can create a draft of their assistants. The organizers will provide course materials and slide deck that participants will be able to utilize to continue development of their custom GPT. The paid subscription to ChatGPT Plus is not required to participate in this workshop, just for trying out personal GPTs during it.
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
Phenomics assisted breeding in crop improvement
As the population is increasing and will reach about 9 billion upto 2050. Also due to climate change, it is difficult to meet the food requirement of such a large population. Facing the challenges presented by resource shortages, climate
change, and increasing global population, crop yield and quality need to be improved in a sustainable way over the coming decades. Genetic improvement by breeding is the best way to increase crop productivity. With the rapid progression of functional
genomics, an increasing number of crop genomes have been sequenced and dozens of genes influencing key agronomic traits have been identified. However, current genome sequence information has not been adequately exploited for understanding
the complex characteristics of multiple gene, owing to a lack of crop phenotypic data. Efficient, automatic, and accurate technologies and platforms that can capture phenotypic data that can
be linked to genomics information for crop improvement at all growth stages have become as important as genotyping. Thus,
high-throughput phenotyping has become the major bottleneck restricting crop breeding. Plant phenomics has been defined as the high-throughput, accurate acquisition and analysis of multi-dimensional phenotypes
during crop growing stages at the organism level, including the cell, tissue, organ, individual plant, plot, and field levels. With the rapid development of novel sensors, imaging technology,
and analysis methods, numerous infrastructure platforms have been developed for phenotyping.
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Nucleic Acid Hybridization.pptx
- 2. CONTENT Hybridization Introduction Southern Hybridization Blotting (Transfer Of DNA On A Membrane) Application DNA Fingerprinting DNA Typing
- 3. Introduction-Hybridization The capacity of denatured DNA to reanneal under appropriate ionic strength and temperature is known as Hybridization. In case of a mixture of DNA molecules, hybridization leads to formation of hybrid molecules. Often a DNA probe is used in hybridization for detecting or finding a particular DNA sequence from a mixture of DNA molecules. Hybridization may be done with DNA fragments separated by electrophoresis, dot blot: DNA spots on a membrane or colony blot: hybridization with bacterial colonies containing particular rDNA.
- 4. Southern Hybridization A method of DNA hybridization established by Ed Southern in 1975. Genomic DNA from an organism is restriction digested and separated by electrophoresis. The electropherosed DNA is transferred on to a membrane. The membrane bound DNA is denatured using an alkali. A DNA probe is radiolabeled and added to the denatured DNA in presence of appropriate buffer for hybridization. After sufficient time for hybridization the membrane is exposed to X-ray film.
- 5. The X-ray film shows signals of hybridization: where the DNA probe has complementary structure on the transferred DNA. The resulting X-ray film is known as an Autoradiogram and the process Autoradiography.
- 7. Blotting (transfer of DNA on a membrane)
- 8. Application Of Southern Blotting Identification of a single gene in a pool of DNA fragments. Gene mapping. Analysis of genetic patterns of dna. Detection of specific dna sequences in a genome. Study of gene deletions, duplications, and mutations that cause various diseases.
- 9. DNA fingerprinting Using the method of DNA hybridization with an appropriate DNA probe to identify individuals as done with fingerprinting. The method was first done by Sir Alec Jeffreys in 1985. Any biological material such as a drop of blood, saliva, semen, and any body part such as bones, tissue, skull, teeth, hair with root etc found at the scene of crime is used as source DNA. DNA probe used for DNA fingerprinting may be a VNTR or minisatellite DNA. The probe is labeled and used to hybridize the source DNA. An autoradiogram is generated that show different band patterns for different individuals.
- 10. Step By Step Procedure Of Fingerprinting
- 11. DNA Fingerprinting Assay Sample
- 12. DNA Typing DNA typing is a procedure wherein DNA extracted from a biological sample obtained from an individual is analyzed. The DNA is processed to generate a pattern for each person that is generally termed as a ' DNA profile'. This profile is unique for each person excepting that derived from identical twins.
- 13. THANK YOU