Viva DNA Extraction
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1. The document describes a study conducted by students to extract DNA from onion cells and chicken liver cells using a conventional method. 2. The methodology involved preparing the onion and chicken liver samples, then grinding or blending the samples with a salt, water and detergent solution. The mixtures were then filtered, heated, and poured into ethanol to isolate the DNA strands. 3. Under a microscope, the students were able to observe DNA strands extracted from both the onion and chicken liver cells using this simple, low-cost conventional method without hazardous chemicals.
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- Chromosomes come in different shapes depending on the location of the centromere. They also have two arms labeled p and q.
- Humans have 46 chromosomes in their somatic cells. The number varies between species but is consistent within a species.
- Chromosomes contain DNA that is tightly packaged and organized through proteins like histones to fit inside the nucleus. Nucleosomes and chromatin compaction allow for the dense packing of DNA.
Eukaryotic chromosome
Eukaryotic genomes are organized into chromatin and packaged at successive levels. DNA is wrapped around histone proteins to form nucleosomes, which are further packaged into higher-order structures like the 30nm fiber and loop domains. These domains are attached to scaffolding proteins, forming interphase chromosome territories and mitotic chromosomes. Precise packaging allows for gene expression control and proper chromosome segregation during cell division. Telomeres and histone modifications also influence chromatin structure and gene regulation.
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3) Precipitating the DNA out of solution by adding alcohol, since DNA is insoluble in alcohol.
4) Isolating the DNA strands using an inoculation loop and transferring them to a small tube for observation.
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- Blending breaks open the cell walls and membranes to release the DNA inside. This physically separates the DNA from other cell contents.
- Salt helps strip away proteins that are attached to the DNA. The positive and negative charges on salt ions disrupt the electrostatic interactions between DNA and proteins.
- Detergent works similarly to salt by disrupting membranes and "poking holes" in them. This allows the contents of the cell to be released. Detergents have molecules with both water-loving and water-fearing parts, allowing them to penetrate and disrupt membranes.
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The document discusses the human genome project, which aimed to sequence the entire human genome and identify all human genes. It provides background on the human genome, describing its size, number of genes, and chromosomes. It details the goals and milestones of the human genome project from 1986 to 2003. Vectors like yeast artificial chromosomes and bacterial artificial chromosomes were used to clone large fragments of DNA for sequencing.
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1. Genetics plays an important role in medicine through studies of inheritance patterns, gene mapping, analysis of disease mechanisms, and diagnosis/treatment of genetic diseases like gene therapy.
2. DNA isolation involves extracting DNA from samples and separating it from other cell components. It is used for scientific research, medicine like outbreak tracing, and forensic science like identification. Various methods disrupt cells, remove proteins, and recover DNA.
3. DNA purification removes contaminants and avoids DNA degradation. Key steps are cell lysis, contaminant removal through various separation techniques, and DNA concentration. Evaluation assesses concentration, purity through absorbance ratios, and degradation using gel electrophoresis.
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DNA fingerprinting is a technique used to identify individuals by analyzing genetic variations in DNA sequences from samples. It involves 7 steps: extracting DNA from samples, cutting DNA into fragments using restriction enzymes, separating fragments by size using gel electrophoresis, transferring fragments to paper, using radioactive probes to identify specific fragments, detecting hybridized fragments through autoradiography, and comparing fragment patterns to determine genetic matches or exclusions. DNA fingerprinting is used for forensic identification, paternity testing, immigration disputes, and missing persons investigations by comparing an individual's unique DNA profile to other samples.
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DNA fingerprinting is a method of identifying individuals based on unique characteristics in their DNA. Alec Jeffreys developed this method in the 1980s to solve criminal cases using DNA profiling techniques. Modern DNA profiling uses short tandem repeats that are variable between individuals and can be used to determine biological relationships in forensic investigations and parental testing. DNA profiling analysis involves extracting DNA from samples, amplifying targeted regions, separating the fragments by size, and comparing the profiles to determine matches or exclusions.
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This document discusses chromosomes and their structure. Some key points:
- Chromosomes are thread-like structures found in the nucleus that contain DNA. They can be seen during cell division.
- Chromosomes come in different shapes depending on the location of the centromere. They also have two arms labeled p and q.
- Humans have 46 chromosomes in their somatic cells. The number varies between species but is consistent within a species.
- Chromosomes contain DNA that is tightly packaged and organized through proteins like histones to fit inside the nucleus. Nucleosomes and chromatin compaction allow for the dense packing of DNA.
Eukaryotic chromosome
Eukaryotic genomes are organized into chromatin and packaged at successive levels. DNA is wrapped around histone proteins to form nucleosomes, which are further packaged into higher-order structures like the 30nm fiber and loop domains. These domains are attached to scaffolding proteins, forming interphase chromosome territories and mitotic chromosomes. Precise packaging allows for gene expression control and proper chromosome segregation during cell division. Telomeres and histone modifications also influence chromatin structure and gene regulation.
Nucleic acid hybridization
Nucleic acid hybridization is a technique where single-stranded nucleic acid molecules form double-stranded molecules through hydrogen bonding between complementary base sequences. This process can identify specific DNA or RNA sequences through the use of labeled probes. There are different types of hybridization including Southern blot, which uses probes to detect complementary DNA sequences separated by electrophoresis; Northern blot, which detects RNA sequences; and colony hybridization, which isolates plasmids containing a particular sequence.
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DNA fingerprinting is a technique used to identify individuals using samples of their DNA. It was invented by Alex Jeffreys in 1985 and involves isolating DNA, digesting it with restriction enzymes, separating the fragments electrophoretically, transferring them to a membrane, probing them, and developing an autoradiograph to visualize the unique pattern of fragments for each individual. DNA fingerprinting has applications in diagnosing inherited disorders, crime investigation by comparing DNA profiles from crime scenes to suspects, determining parentage in cases, and personal identification such as for military casualties.
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In eukaryotic cells, DNA is packaged through interactions with histone proteins to form nucleosomes, which are beads of DNA wound around histone octamers. Nucleosomes further condense into chromatin, and additional non-histone proteins compact chromatin into higher-order structures. In prokaryotes, DNA is loosely organized into loops associated with proteins in the nucleoid region, which lacks a membrane-bound nucleus. Eukaryotic chromatin also exists in two forms - loosely coiled euchromatin containing active genes, and tightly packed heterochromatin containing inactive regions.
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- Salt helps strip away proteins that are attached to the DNA. The positive and negative charges on salt ions disrupt the electrostatic interactions between DNA and proteins.
- Detergent works similarly to salt by disrupting membranes and "poking holes" in them. This allows the contents of the cell to be released. Detergents have molecules with both water-loving and water-fearing parts, allowing them to penetrate and disrupt membranes.
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ေန႔စဥ္ၿငိမ္းေရးသတင္းစဥ္ ၊ မတ္လ (၂၀) ရက္ေန႔ ၊ ၂၀၁၇ ခုႏွစ္။ ခ်င္းေအဂ်င္စီ
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The document describes techniques for extracting, purifying, and quantifying DNA. It discusses that DNA can be extracted from biological materials using various chemical and physical methods. Common extraction methods mentioned include phenol-chloroform extraction, proteinase K digestion, salting out, and silica adsorption. Purification techniques help remove contaminants, and quantification is usually done using UV spectrophotometry, agarose gel electrophoresis, or the NanoDrop system to determine DNA concentration and purity.
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DNA must be purified for various applications. Key steps in DNA purification include cell lysis, removal of contaminants like proteins and RNA, and concentrating the purified DNA. Common methods for purification involve organic extraction using phenol-chloroform, binding DNA to a solid support like silica or magnetic beads, and ethanol precipitation. Evaluation of purified DNA includes measuring concentration using UV absorbance and assessing purity based on absorbance ratios. Purifying high quality DNA is essential for downstream applications in research, medicine, and forensics.
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1. Genetics plays an important role in medicine through studies of inheritance patterns, gene mapping, analysis of disease mechanisms, and diagnosis/treatment of genetic diseases like gene therapy.
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1. Genetics plays an important role in medicine through studies of inheritance patterns, gene mapping, analysis of disease mechanisms, and diagnosis/treatment of genetic diseases like gene therapy.
2. DNA isolation involves extracting DNA from samples and separating it from other cell components. It is used for scientific research, medicine like outbreak tracing, and forensic science like identification. Various methods disrupt cells, remove proteins, and recover DNA.
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- 1. EXTRACTION OF DNA FROM ONION CELL AND CHICKEN LIVER BY: NUR SHUHADA NABILLA BINTI MOHD YUNUS NUR LAILI BINTI YEOP IDRIS RAIHAN IZZATI BINTI ROSANIZAN NUR ATIELIA BINTI PRESHAHDIN NURUL AFIFA BINTI ABDUL RAHMAN SUPERVISOR: DR. ZAKIAH BINTI NOR HASHIM
- 2. PROBLEM STATEMENT 1.Can we observed DNA strand with naked eyes? 2. Can we obtained the DNA strand by using conventional method?
- 3. HYPOTHESIS Conventional method is more reliable compared to various commercial offerings
- 4. INTRODUCTION DNA : Deoxyribonucleic acid, self replicating material present in nearly all living organisms, as the main constituents of chromosomes and carrier of genetic information. DNA Extraction: Removal of Deoxyribonucleic acid (DNA) from cells of sample. General Structure: • Double helix (composed of 2 chain) • Strands of nucleotide held together by hydrogen bond
- 5. Background of study DNA is present in every cell of plants and animals The long and thick fibers of DNA Can be extracted using common, everyday materials Extraction buffer
- 6. OBJECTIVES 1. To learn the steps of preparing chicken liver and onion cell for DNA extraction. 2. To understand the general structure of these living tissues. 3. To extract DNA from cells at a simple, reasonable cost and no hazardous chemicals are involved by using conventional method.
- 7. SIGNIFICANCE OF STUDY 1. Can gain foundation knowledge for lesson in biology, evolution, and biotechnology and health technology. 2. Conventional method able to produce high yield, less time consuming and reliability of the analysis should be maximal, while minimizing the risk of contamination
- 9. Titles References Methods DNA extraction Karen, 1999 • “DNA Extraction” are to isolate DNA from plant and animal cells. • Serves as a spectacular example of the complexity of biochemical structure DNA extraction from animal and plant tissues Karen, 1999 • Plant genomic DNA is more difficult to extract. • The metabolites present in the plant cell may interfere with genomic DNA extraction. Methods of DNA extraction Shepherd, 2002 • Several commercial kits are available that allow rapid and reliable isolation of total DNA from a wide of variety of plant species and tissues. Estimation of DNA quantity K.K Vinod, 2004 • Most DNA extracts are not reasonably pure and therefore estimates of concentration using spectrophotometric measurements of UV absorption may be misleading.
- 10. METHODOLOGY Preparation of DNA extraction of chicken liver cell Preparation of DNA extraction of onion cell
- 11. Preparation of DNA extraction of onion cell Chopped the onion Poured water, salt and detergent Poured the solution into the blender Heat the mixtureFiltered heated mixture Poured the sample into ethanol
- 12. Preparation of DNA extraction of chicken liver cell Grind the chicken liver Poured the water, salt and detergent Stir the mixture well Filtered the mixturePoured mixture into ethanol The DNA strand observed
- 13. Chicken liver cell DNA strand result Under 10x magnification
- 14. Onion cell DNA strand results
- 15. CONCLUSION 1. We are able to learnt the steps of preparing chicken liver and onion cell for DNA extraction. 2. The general structure of these living tissues were well defined. 3. We are able to extract DNA from cells at a simple, reasonable cost and no hazardous chemicals are involved by using conventional method.
- 16. RECOMMENDATION 1. Spectrophotometer To measure the purity of the DNA extraction 2. Agarose gel electrophoresis Separation of DNA fragments according to size, based on movement, through a gel medium when an electric field is applied.
- 17. The End