This lectureis about DNA extraction from whole Blood presented by Tuba nafees she is msc graduate in Biotechnology from University of Karachi, Sindh Pakistan.
lecture video is also there in youtube link:
https://www.youtube.com/watch?v=cGr__SuqYgY&t=409s
Basics of DNA isolation, What is chemistry behind it. Presently the laboratory of animal science department ,Göttingen university using this technique for dna isolation in pig blood sample.
This presentation is about DNA fingerprinting, a brief description is given about its principle, working, technique and its application with a example.
Techniques of DNA Extraction, Purification and QuantificationBHUMI GAMETI
Introduction
The overall process…
Uses of isolated genomic DNA
Extraction of DNA from plant material
Components of DNA extraction solutions
Cell Lysis or Cell disruption :
Purification of DNA
CTAB Method
Phenol–chloroform extraction
PROTEINASE K
Salting out
Silica adsorption method
Magnetic beads
FTA Paper
Nucleic acid quantification
Agarose Gel Electrophoresis
UV spectroscopy
DNA quantification using NanoDrop
There are 'n' number of DNA isolation methods depending on the sample type, final use of DNA product, etc. This presentation gives an overall idea about different methods of DNA isolation in a simplified way.
Basics of DNA isolation, What is chemistry behind it. Presently the laboratory of animal science department ,Göttingen university using this technique for dna isolation in pig blood sample.
This presentation is about DNA fingerprinting, a brief description is given about its principle, working, technique and its application with a example.
Techniques of DNA Extraction, Purification and QuantificationBHUMI GAMETI
Introduction
The overall process…
Uses of isolated genomic DNA
Extraction of DNA from plant material
Components of DNA extraction solutions
Cell Lysis or Cell disruption :
Purification of DNA
CTAB Method
Phenol–chloroform extraction
PROTEINASE K
Salting out
Silica adsorption method
Magnetic beads
FTA Paper
Nucleic acid quantification
Agarose Gel Electrophoresis
UV spectroscopy
DNA quantification using NanoDrop
There are 'n' number of DNA isolation methods depending on the sample type, final use of DNA product, etc. This presentation gives an overall idea about different methods of DNA isolation in a simplified way.
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
this section helps students how to quanify the isolated DNA by spectrophotometer. specially life life science fields such as biotechnology, biology, and medical laboratory
DNA fingerprinting is a research facility procedure used to set up a connection between natural proof and a suspect in a criminal examination. A DNA test taken from a wrongdoing scene is contrasted and a DNA test from a suspect. On the off chance that the two DNA profiles are a match, at that point the proof originated from that suspect. On the other hand, on the off chance that the two DNA profiles don't coordinate, at that point the proof can't have originated from the suspect. DNA fingerprinting is likewise used to build up paternity.
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes to investigate the presence of small, submicroscopic chromosomal changes that are beyond the resolution of karyotype analysis.
This PowerPoint presentation explain the concept,process and application of Fluorescence insitu hybridization.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
this section helps students how to quanify the isolated DNA by spectrophotometer. specially life life science fields such as biotechnology, biology, and medical laboratory
DNA fingerprinting is a research facility procedure used to set up a connection between natural proof and a suspect in a criminal examination. A DNA test taken from a wrongdoing scene is contrasted and a DNA test from a suspect. On the off chance that the two DNA profiles are a match, at that point the proof originated from that suspect. On the other hand, on the off chance that the two DNA profiles don't coordinate, at that point the proof can't have originated from the suspect. DNA fingerprinting is likewise used to build up paternity.
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes to investigate the presence of small, submicroscopic chromosomal changes that are beyond the resolution of karyotype analysis.
This PowerPoint presentation explain the concept,process and application of Fluorescence insitu hybridization.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Lab 23 DNA Extraction and PurificationIsolation and purific.docxDIPESH30
Lab 2/3: DNA Extraction and Purification
Isolation and purification of nucleic acids is the most fundamental procedure in molecular biology. There are three basic steps involved:
1. Lyse (break open) the cells (and nuclei in eukaryotes) to release the DNA
2. Remove contaminants (proteins, lipids, carbohydrates, salts)
3. Preserve the integrity of the DNA (prevent degradation and shearing)
Step 1 can be accomplished in a number of ways, such as mechanical disruption (grinding, mincing), protein denaturation (detergents), and protein degradation (via proteases). These can be used singly or in combination depending on the type of biological sample you are starting with. Grinding the samples provides more surface area for the denaturants/proteases to interact with the cellular proteins, thus speeding up the denaturation process. We used liquid nitrogen (N2) and protein degradation (Proteinase K) in lab 2. Various salts are included in a cell lysis solution to stabilize the DNA by providing positive ions which insert between the negatively charged phosphates in the DNA backbone (creating a “salt bridge”). Buffers (such as Tris) also help to preserve DNA integrity by maintaining a neutral pH.
Once the cells have been lysed, contaminating proteins, lipids, etc. must be separated from the DNA. A widely used and efficient way to remove proteins from nucleic acids solutions is to extract with a 1:1 mixture of phenol and chloroform (CHCl3). Phenol and CHCl3 are both hydrophobic organic solvents that unfold proteins. When mixed with an aqueous DNA/protein solution and then centrifuged, the denatured proteins are selectively partitioned into the denser organic phase, while the DNA (plus RNA and salt) remains in the aqueous phase. This procedure takes advantage of the fact that deproteinization is more efficient when two different organic solvents are used instead of one. Additionally, chloroform removes any lingering traces of phenol from the nucleic acid preparation (which would interfere with later applications). Since the aqueous phase contains RNA and salt in addition to the DNA, phenol:CHCl3 extraction is followed by ethanol (EtOH) precipitation. DNA (a polar molecule) is soluble in water (also polar) because the water molecules intercalate into the phosphate backbone of the DNA and thus maintain it in a soluble state, but DNA is insoluble in 95% EtOH (nonpolar). Water molecules have a higher affinity for the EtOH than the DNA, so when you add EtOH and salt [10 M ammonium acetate (NH4Ac); pH 5.2], Na+ ions replace water in the DNA backbone, essentially removing the water molecules, and the DNA is forced out of solution (precipitates). After precipitating with 95% EtOH, the DNA is “washed” in 70% EtOH to remove the salt. Since 70% EtOH contains 30% water, the salt, having a greater affinity for the water than the DNA, remains in the EtOH, and the DNA is forced out.
The final step in the purification process is to preserve the DNA in a sta ...
DNA Fingerprinting & its techniques by Shiv Kalia (M.Pharma in Analytical Che...Shiv Kalia
DNA fingerprinting and below mention content widely cover in this presentation
History & Introduction of DNA fingerprinting
How was the first DNA fingerprint produced?
Types of DNA Based Markers
Polymerase Chain Reaction (PCR)
PCR based Methodology of DNA fingerprinting
Electrophoresis
Utility of DNA Based Markers
Various DNA Fingerprinting Techniques Advantages & Disadvantages
Authentication of Various Ayurvedic Herbs by DNA Fingerprinting
Advantages of DNA fingerprinting in Plants
Disadvantages of DNA fingerprinting in Plants
CONCLUSION
Sustainable Practices in Event ManagementSindhBiotech
The following presentation is presented by our intern, Lucky Qindeel Panhwar, from Phase 1 of the ongoing SBA Internship Program.
In the era of acute pollution and threatening social and environmental conditions, the utmost importance must be given to finding and applying 'green solutions'. Because such a basis will provide both ecological and economic sustainability while ensuring good social practices.
The important is that "Sustainable practices must be in action, not just in the mind".
Launching an event successfully hinges on thoughtful planning and execution. The
Implementation of strategies that prioritize eco-friendly catering, sustainable venues, and environmentally conscious promotions, is key to effective Sustainable Event Management.
For Youtube: https://youtu.be/ybPsyld3wlM?si=h9NM3PQfoyoWZKJQ
#GreenMarketing #SustainableEventsforSustainableSociety #EcoFriendlyApproaches #GreenCatering
#KarachiGreenMarathon
Potentials of Microfluids in Life Sciences: A Lab on a ChipSindhBiotech
Today marks the death anniversary of Mr. Werner Jacobi, the pioneering figure in microfluidics. While his initial focus was on microelectronics and semiconductor technology, he carved out a distinct path for microfluidics, diverging from his previous pursuits. His visionary approach continues to shape the trajectory of microfluidics research and its diverse applications in science and technology.
This lecture is presented by our team member and volunteer, Bushra Umer, from Volunteer Batch-V, 2023.
The lecture emphasizes the adoption of the "lab-on-a-chip" approach to ensure cost-effective and real-time experiments, underscoring the importance of developing such innovative techniques in today's modern arena.
For Video: https://youtu.be/ZF-c6k-F1RI
#wernerjacobi
#biotechnology
#technology
#lifesciences
#microfluidics
How Bio-inspired Plastic Outperforms Traditional Plastic: A Comparative AnalysisSindhBiotech
This presentation is brought to you by Bushra Umer, our team member and volunteer from Batch - V, 2023.
It highlights the significance of bioplastics, which outshine traditional plastics in sustainability and performance, paving the way for a cleaner, greener, and healthier planet.
Considering the ongoing environmental crisis, there is an urgent necessity to embrace such sustainable solutions to rescue Mother Earth from environmental calamities.
#Bioplastics #SustainableLiving #GreenRevolution #WorldEarthDay
Youtube Link: https://youtu.be/LFpYs_4N_1k
Common Genetic Disorders Prevailing in PakistanSindhBiotech
This Lecture is presented by our 2k23 volunteer and Content Presenter Zargull Sadein , she is from Karachi, Pakistan, and she is covering "Common Genetic Disorders Prevailing in Pakistan".
Youtube: https://youtu.be/16scqaB1yjk
The Revolutionary Progress of Artificial Inteligence (AI) in Health CareSindhBiotech
This Lecture is presented by our 2k23 volunteer Hina Nawaz, she is from Karachi, Pakistan, and she is covering "The Revolutionary Progress of Artificial Inteligence (AI) in Health Care".
Youtube: https://youtu.be/vhJRCj5ZgJc
Nanobots: Lecture on the Artificial BloodSindhBiotech
This Lecture is presented by our volunteer Laraib Elahi, she is from Karachi, Pakistan, and she is covering "Nanobots: The Artificial Blood. "
For video: https://youtu.be/TsUgvOh6tOA
Decoding the Monkeypox Virus : From Discovery to PreventionSindhBiotech
This lecture is presented by our volunteer Sajid Ali Shah, he is from Islamabad, Pakistan, and he is covering the topic Decoding the Monkeypox Virus : From Discovery to Prevention.
For video: https://youtu.be/-RT2UvTerBc
This Lecture is presented by our volunteer Abrar Hussain, he is from Karachi Pakistan, and he is covering The escence of research.
Youtube: https://youtu.be/CB0CY70K2go
The role of CRISPR CAS-9 in the treatment of HIVSindhBiotech
This lecture is presented by our volunteer Sajid Ali Shah, he is from Islamabad, Pakistan, and he is covering The role of CRISPR CAS-9 in the treatment of HIV.
for video: https://youtu.be/c0gGdzKUACI
Developing Therapeutic Approaches For Emerging Viral DiseasesSindhBiotech
This lecture is presented by our volunteer Zargul sadein, she is from Karachi, Pakistan, and she is covering "Developing Therapeutic Approaches For Emerging Viral Diseases."
for video: https://www.youtube.com/watch?v=DcI89jpjbxc
unwinding the potentials of stem cellsSindhBiotech
This lecture is presented by our volunteer Javeria Khan, she is from Karachi, Pakistan, and she is covering "unwinding the potentials of stem cells"
for video: https://youtu.be/GDC3RKwIACY
Designer babies : A Health Wonder Or an Ethical ErrorSindhBiotech
This lecture is presented by our volunteer Laraib Elahi, she is from Karachi, Pakistan, and she is covering "Designer babies : A Health Wonder Or an Ethical Error. "
For video: https://youtu.be/YGZQGL2QHuU
Personalized Medicines - Enhancers of Life's Quality and Their Future SindhBiotech
This lecture is presented by our volunteer Bushra Umer, she is from Karachi, Pakistan, and she is covering "Personalized Medicines - Enhancers of Life's Quality and Their Future ".
For video: https://www.youtube.com/watch?v=BSrgJaBYuxg
Conceptual Understanding of Monoclonal Bodies Production via Hybirdoma Techno...SindhBiotech
This video is presented by our volunteer Mehwish Khan, she is from Karachi, Pakistan, and she is covering "Monoclonal Bodies Production via Hybirdoma Technology"
for video: https://youtu.be/NilP7HIALvY
#Hybirdoma #antibody #monoclonalantibodies #biology #physiology #health #polyclonalantibody
Microbiota and Gut-Brain Axis in HealthSindhBiotech
This lecture is presented by our volunteer Hina Nawaz, she is from Karachi, Pakistan, and she is covering "Microbiota and Gut-Brain Axis in Health"
For video: https://youtu.be/W2hfl5_FaF8
Biosensors in Environmental MonitoringSindhBiotech
This lecture is presented by our volunteer Bushra Umer, she is from Karachi, Pakistan, and she is covering "Biosensors in Environmental Monitoring"
For video: https://youtu.be/DoO2Aw7bRrk
Regulation and Integration of MetabolismSindhBiotech
This lecture is presented by our volunteer Hina Nawaz, she is from Karachi, Pakistan, and she is covering "Regulation and Integration of Metabolism"
for video: https://youtu.be/D2k-_f28TQA
Dengue Virus: Genomic Insights, Pathogenic Mechanisms, and Therapeutic Approa...SindhBiotech
This lecture is presented by our volunteer Sajid Ali Shah, he is from Islamabad, Pakistan, and he is covering Dengue Virus: Genomic Insights, Pathogenic Mechanisms, and Therapeutic Approaches
for video: https://youtu.be/whrkkKR-NSY
#denguevirus #virology #virologist #genomics #covid19 #virus #pathogen #pathology #immunology
Gene environment interaction and its impact upon on human healthSindhBiotech
This lecture is presented by our volunteer Zargull Sadein, she is from Karachi Pakistan, and she is covering Gene environment interaction and its impact upon on human health.
for video: https://youtu.be/97F1tR9jj5k
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.
Richard's aventures in two entangled wonderlandsRichard 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.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
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.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
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.
2. PRINCIPLE OF DNA EXTRACTION
1. CELL LYSIS
2. SEPARATION OF DNA FROM
CELL DEBRIS
3. PURIFICATION OF DNA
2
3. METHODS OF DNA EXTRACTION
1. Phenol-chloroform extraction
2. Magnetic bead separation
3. Precipitation chemistry (or “Salting out”).
When the ionic strength of a protein solution is increased
by adding salt, the solubility decreases, and protein
precipitates
Results in little-to-no contamination, gives high DNA yields
and is cost-effective.
ADVANTAGES
3
Salt
molecules Protein
molecules
5. DNA EXTRACTION BY (Salting Out/
Precipitation Chemistry)
1. Cell lysis (To expose
DNA)
• Buffer (To maintain pH),
Detergents (To break lipids)
• PROTEASE (To break proteins)
• RNase (To degrade RNA)
CELL LYSIS
BUFFER
Use of high concentration of NaCl ( 6
M). After protein precipitation DNA
floats freely in the supernatant and is
separated from clumped cell debris
2. Precipitation of cell debris
4. Precipitation/Purification of
DNA
Isoproanol/ Ethanol precipitation,
minicolumn binding.
ADDIOTIONAL
REAGENTS
Cell lysis buffer uses NaCl (in low conc) to
precipitate cell debris ( detergents,
broken proteins, RNA , lipids etc)
3. Precipitation of Protein
5
7. METHODOLOGY
EDTA
EDTA acts as an
“Anticoagulant” ( It
prevents blood
clotting)and is the
best choice for blood
DNA extraction than
heparin and sodium
citrate.
SAMPLE STORAGE
Within 3 days =
Refrigerate at 4 ͦC
For longer use =
Store at -80 ͦC
SAMPLE PREPARATION
7
8. Blood sample
invert mixing
Cell lysis
Buffer
Centrifugation at 4000 rpm
for 10 min
Discard
supernatant
in a new
microfuge
tube
Addition of
SDS
8
Cell debris
9. Incubate at
50 °C for 30
min
Addition of
NaCl
Collection of
Supernatant
Centrifugation at 4500
rpm for 15 min
9
DNA
Precipitation of
Proteins
10. Addition of 95 %
ice- cold
isopropanol
Centrifugation at
12000 rpm for 4 min 10
11. Wash the pellet
with 70 %
ethanol
Centrifugation at
12000 rpm for 4 min
Air dry and
Resuspend in TE
buffer ( pH 8 )
/Pure water
11
12. DETECTION OF DNA
BY GEL ELECTROPHORESIS
• The gel consists of a permeable matrix, a
bit like a sieve, through which molecules
can travel when an electric current is
passed across it.
• Smaller molecules migrate through the gel
more faster than larger fragments
• To visualise the DNA, the gel is stained
with a fluorescent dye (ethidium bromide)
that binds to the DNA, and is placed on an
ultraviolet transilluminator which will
show up the stained DNA as orange bands.
• The length of the DNA fragments is
compared to a marker containing
fragments of known length.
12
DNA
MARKER
Sample DNA
BANDS
Smaller
fragment
Larger fragment
13. PURITY OF DNA
• Take absorbance of DNA at two different
wavelengths i.e. 260nm and 280nm and then
finding their ratio. DNA absorbs UV light both at
260 and 280 nm while proteins absorbs UV at
only 280nm.
• Pure DNA with no protein contamination will give
a ratio of 1.8 at 260/280. On the other hand, a
DNA with protein impurities will give lower value
than 1.8.
13
16. REFERENCES
• Sugna et al. (2014). DNA Isolation from Human Whole Blood
Samples by Non-Enzymatic Salting Out Method.
International Journal of Pharmacy and Pharmaceutical
Sciences, Vol 6, 198-199.
• Shahid. (2019). Genomic DNA Extraction – Principle, Steps
and Functions of Reagents. How Biotech. Retrieved from
http://howbiotech.com/genomic-dna-extraction-principle-
steps-and-functions-of-reagents
• Miller, S.A., Dykes, D.D., Polesky, H.F. (1998). A Simple
Salting Out Procedure for Extracting DNA from Human
Nucleated. Nucleic Acids Research,16(3), 1215
• Rice G.DNA Extraction. Educational Resources, Microbial
Life. Montana State University. Retrieved 17 February 2017.
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