Elektroporasyon cihazi
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Electroporation device design, electroporation method
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Culture stage 1 initiation stage
Introduction
Advantages of Micropropagation over the conventional methods
History
Stages of Micropropagation
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Mass selection 21.05.2021
This document discusses mass selection, a method of plant breeding where individual plants are selected based on desirable phenotypes from a mixed population. Seeds from selected plants are then combined to grow the next generation. Mass selection can be used to improve self-pollinating or cross-pollinating crops. It has resulted in improved varieties of crops like bajra, yellow sarson, brown sarson, maize, desi cotton, and castor. While mass selection is simple and applicable to many crops, the improvements may be less than other methods and varieties developed have more genetic variation than pure lines.
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Artificial seeds, also known as synthetic seeds, involve the encapsulation of somatic embryos, shoot buds, or cell aggregates to propagate plants in vitro or ex vivo. They were first introduced in the 1970s and provide advantages like large-scale and low-cost propagation while maintaining genetic uniformity. Successful artificial seeds require an embryo-protective coating containing nutrients to support germination and growth. The coating material, embryo maturity, and encapsulation process can produce either desiccated or hydrated synthetic seeds. Common steps in artificial seed production involve establishing embryogenesis, encapsulating mature embryos, and field planting.
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Introduction
Importance of Protoplast Isolation
Source of Protoplast
Isolation of Protoplast
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Culture of Protoplast
Protoplast Regeneration
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Cybrids
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Somatic cell cloning and dolly ppt.pptx
This document discusses somatic cell cloning. Somatic cells are non-reproductive body cells that can differentiate. Somatic cell cloning involves transferring the nucleus of a somatic cell into an egg cell to produce a cloned embryo. Dolly the sheep was the first cloned mammal, created via somatic cell cloning in 1996. The process of somatic cell cloning and the story of Dolly's creation are described. Ethical issues with cloning like animal welfare, unintended consequences, commercialization, and loss of genetic diversity are also discussed.
Plant Protoplast: Isolation, Purification and Culturing
The document discusses plant protoplast isolation, purification, and culturing. Some key points:
- Protoplasts are plant cells that have had their cell walls removed, leaving just the plasma membrane. They allow for plant cell fusion and regeneration.
- Protoplasts are typically isolated from plant tissues like leaves using enzymatic digestion with cellulase and pectinase. This yields more protoplasts than mechanical methods.
- Isolated protoplasts are purified by centrifugation and washing to remove cell debris. They are then cultured in liquid or solid nutrient media and tested for viability before regeneration.
Protoplast isolation and immobiliz by Dr.U.Srinivasa
The document discusses protoplasts, which are plant cells that have had their cell walls removed, leaving the cell membrane and organelles. It describes methods for isolating protoplasts from plant tissues using either mechanical or enzymatic methods. The enzymatic method uses enzymes like pectinase and cellulase to break down the cell wall. Protoplasts have various applications including isolating cell organelles and studying cell structures. The document also discusses immobilizing enzymes by binding them to inert matrices, which has benefits like reusability and stability. Methods of immobilization include adsorption, covalent binding, and entrapment in gels.
Culture stage 1 initiation stage
Introduction
Advantages of Micropropagation over the conventional methods
History
Stages of Micropropagation
1. Stage 0; Preparative stage
2. Stage 1; Initiation of aseptic cultures
A) Explant
B) Sterilization
C) Browning of medium
Factors affecting initiation stage
Conclusions
References
Somatic ybridization and its application
This document discusses somatic hybridization, which involves fusing plant protoplasts from two different species or varieties to create a hybrid plant. It describes the process of somatic hybridization, including isolating protoplasts, fusing them using spontaneous or induced methods, selecting hybrid cells, and regenerating plants from hybrid callus tissue. The advantages are producing novel hybrids and transferring genes between incompatible species. The limitations include low regeneration rates and viability of fused cells. Somatic hybridization has applications in crop improvement by introducing traits like disease resistance from wild relatives.
Vector less gene transfer
This document discusses various direct or vectorless gene transfer methods for introducing foreign DNA into plant cells without the use of Agrobacterium. These include chemical methods like PEG-mediated transformation, calcium phosphate precipitation, DEAE-dextran, and liposome-mediated transformation. Physical methods include electroporation, microinjection, and gene guns. The procedures, advantages, and disadvantages of some key methods like PEG-mediated transformation, calcium phosphate precipitation, electroporation, microinjection, and fiber-mediated transformation are described in detail. Direct transformation through imbibition of dehydrated plant tissues is also mentioned.
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Terminator technology
Terminator gene technology refers to plants that have been genetically modified to render sterile seeds at harvest.
Genetic use restriction technologies (GURTs) are the name given to experimental methods, described in a series of recent patent applications and providing specific genetic switch mechanisms that restrict the unauthorized use of genetic material (FAO, 2001a) by hampering reproduction (variety-specific V-GURT) or the expression of a trait (trait-specific T-GURT) in a genetically modified (GM) plant.
Micropropagation
Micropropagation is an advanced vegetative propagation technology for producing a large number of transplants in a limited time and space.
STAGES
Stage 0 — Mother Plant Selection:
Stage I — Establishment of Aseptic Culture:
Stage II — Multiplication of shoots:
Stage III — In Vitro Rooting:
Stage IV — Transplantation or Hardening:
APPLICAIONS
M S Medium Preparation
Murashige and Skoog medium was originally formulated in 1962 to optimize tobacco callus growth and study cytokinins. It has since become widely used for micropropagation, organ culture, callus culture, and suspension culture of plant tissues. The medium is a nutrient blend of inorganic salts, vitamins, and amino acids that provides the necessary nutritional requirements for cultured plant cells and tissues to grow. It contains macro and micro nutrients, vitamins, amino acids or other nitrogen supplements, sugars or other organic supplements, solidifying agents, and plant growth regulators.
Mutation & its detection
1. The document discusses mutation and its detection. It defines mutation as heritable changes in the genome excluding those from other organisms.
2. It describes different types of mutations such as spontaneous versus induced, forward versus reverse, nuclear versus cytoplasmic, and more.
3. Methods of detecting mutations in prokaryotes and eukaryotes are described. For prokaryotes, techniques like replica plating and the Ames test are used. For eukaryotes, each individual must be examined for mutant phenotypes.
Somatic hybridization faizy
This document provides an introduction to somatic hybridization, which is the fusion of somatic protoplasts from two different plant species or varieties to create hybrid plants. It discusses the history and development of the technique, including early work isolating protoplasts. The key steps in somatic hybridization are described: isolating protoplasts using either mechanical or enzymatic methods, fusing the protoplasts spontaneously or through induction methods, identifying and selecting hybrid cells, culturing the hybrid cells, and regenerating hybrid plants. Advantages include creating novel hybrids and transferring desirable traits between species, while limitations include low regeneration rates and viability of fused products.
Genetic recombination in_bacteria-jb
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Protein targetting ppt (1)
This document summarizes protein targeting mechanisms in cells. It discusses how signal sequences direct proteins to different organelles like the endoplasmic reticulum, mitochondria, chloroplasts and nucleus. The signal sequence is cleaved after the protein reaches its destination. Glycosylation in the ER plays a key role in targeting lysosomal enzymes. Mitochondrial and chloroplast proteins use a different targeting mechanism after full synthesis. Receptor-mediated endocytosis imports some extracellular proteins by binding to receptors and forming clathrin-coated vesicles.
Anther culture
Anther culture is a technique where anthers are excised from flower buds and cultured on nutrient media. This can produce haploid plantlets through either organogenesis or embryogenesis. The first report of haploid tissue from anther culture was in 1966 in Datura pollen grains. Haploids are useful for plant breeding as they contain only one allele per gene, revealing recessive traits, eliminating lethal genes, and allowing for efficient production of homozygous plants. The protocol involves sterilizing tobacco flower buds, removing anthers, and culturing them on nutrient media. After 3-4 weeks, haploid plantlets emerge from the cultured anthers. Haploids have applications in basic research, mutation studies
totipotency
Totipotency refers to a plant cell's genetic potential to regenerate an entire plant. In plant tissue culture, cells taken from plant parts can form an unorganized mass of cells called callus tissue when placed in a sterile nutrient medium. The cells within callus tissue are totipotent, meaning they have the ability to regenerate into a whole plant under the right conditions and hormone manipulations. Totipotency is important for plant science applications like vegetative propagation of crops, genetic modification of plants, and preservation of endangered plant species through long-term storage of totipotent cells.
Anther culture
Anther culture is an in vitro technique used to produce haploid plants from male gamete cells. Haploid cells contain only one set of chromosomes. The first report of anther culture was in the 1970s, while the first natural occurrence of haploids was observed in 1922 in Datura plants. Anthers containing microspores are cultured on nutrient media supplemented with hormones and sugars. Depending on the species, haploid plants or callus can regenerate from the cultured anthers within 3-8 weeks. Haploid plants are useful for breeding programs as they can be doubled to generate fertile, homozygous plants in one generation.
PROTOPLAST ISOLATION & FUSION
This document discusses protoplast isolation and fusion. It defines a protoplast as a plant, bacterial, or fungal cell that has had its cell wall removed, leaving the plasma membrane intact. The document outlines the history of protoplast isolation, from the first isolation in 1892 to the use of commercial enzyme mixtures in the 1960s. It also describes how protoplasts can be fused using polyethylene glycol or other fusogens to create somatic hybrids for plant breeding purposes.
Special type of chromosomes
This document discusses two special types of chromosomes: polytene chromosomes and lampbrush chromosomes. Polytene chromosomes are found in salivary glands and are formed through repeated DNA replication without cell division, resulting in fused sister chromatids. Lampbrush chromosomes are found in growing oocytes of birds and some animals, and appear during meiosis with large chromatin loops and chromomeres that give them a lampbrush-like appearance used for active gene transcription. Both special types of chromosomes are much larger than normal chromosomes and are involved in high levels of gene expression or RNA transcription.
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Protoplast Culture
A presentation covering the process of protoplast culture including protoplast isolation, protoplast fusion, culture of protoplast, its application, factors affecting protoplast culture and the future of protoplasts.
Protoplast fusion
Protoplast fusion involves removing the cell walls of plant cells through enzymatic or mechanical means to create naked protoplasts. These protoplasts can then be fused using chemicals, electricity, or other methods. This allows the cytoplasms and sometimes nuclei of different plant cells to merge, creating hybrid cells. Successful fusion can generate hybrid plants through regeneration of cell walls and tissues. Protoplast fusion overcomes sexual incompatibility and is used to introduce traits like disease resistance between species. It remains a technically challenging process with limitations like genetic instability and uncertain expression of transferred traits.
Suspension Culture and Single Cell Cultures, Culturing methods, maintenance a...
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Generally, suspension culture is a one stop technology to produce secondary metabolites on a large scale in-vitro, irrespective of the climatic condition or nutrient availability (as required in field plants).
In this presentation, we will see the importance of suspension culture, culturing methods and it's application (mostly with respect to plants) and also focus on what exactly is a single cell culture.
Development in arabidopsis
Introduction And Classification
Anatomy Of Flower
Life Cycle Of Arabidopsis
Early Flower Development
Embryogenesis-
A. Formation Of Microspores
B. Formation Of Megaspores
Embryonic Development Starts By Establishing A Root-shoot Axis And Then Halts Inside The Seed
Arabidopsis Genome Is Rich In Developmental Control Genes.
Control Of Carpel & Fruit Development
Arabidopsis Thaliana A Model Plant
Conclusion
References
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Somatic ybridization and its application
This document discusses somatic hybridization, which involves fusing plant protoplasts from two different species or varieties to create a hybrid plant. It describes the process of somatic hybridization, including isolating protoplasts, fusing them using spontaneous or induced methods, selecting hybrid cells, and regenerating plants from hybrid callus tissue. The advantages are producing novel hybrids and transferring genes between incompatible species. The limitations include low regeneration rates and viability of fused cells. Somatic hybridization has applications in crop improvement by introducing traits like disease resistance from wild relatives.
Vector less gene transfer
This document discusses various direct or vectorless gene transfer methods for introducing foreign DNA into plant cells without the use of Agrobacterium. These include chemical methods like PEG-mediated transformation, calcium phosphate precipitation, DEAE-dextran, and liposome-mediated transformation. Physical methods include electroporation, microinjection, and gene guns. The procedures, advantages, and disadvantages of some key methods like PEG-mediated transformation, calcium phosphate precipitation, electroporation, microinjection, and fiber-mediated transformation are described in detail. Direct transformation through imbibition of dehydrated plant tissues is also mentioned.
E. coli as a model organism in research
E. coli is a model organism commonly used in research due to its small genome, rapid reproduction rate, and ability to be easily manipulated and engineered. As a prokaryotic and unicellular organism, E. coli is useful for research because it can be grown and studied without ethical concerns. Its simple nutrient needs allow it to adapt to different environments and be cultured for multiple generations in a short period of time to advance scientific understanding.
Terminator technology
Terminator gene technology refers to plants that have been genetically modified to render sterile seeds at harvest.
Genetic use restriction technologies (GURTs) are the name given to experimental methods, described in a series of recent patent applications and providing specific genetic switch mechanisms that restrict the unauthorized use of genetic material (FAO, 2001a) by hampering reproduction (variety-specific V-GURT) or the expression of a trait (trait-specific T-GURT) in a genetically modified (GM) plant.
Micropropagation
Micropropagation is an advanced vegetative propagation technology for producing a large number of transplants in a limited time and space.
STAGES
Stage 0 — Mother Plant Selection:
Stage I — Establishment of Aseptic Culture:
Stage II — Multiplication of shoots:
Stage III — In Vitro Rooting:
Stage IV — Transplantation or Hardening:
APPLICAIONS
M S Medium Preparation
Murashige and Skoog medium was originally formulated in 1962 to optimize tobacco callus growth and study cytokinins. It has since become widely used for micropropagation, organ culture, callus culture, and suspension culture of plant tissues. The medium is a nutrient blend of inorganic salts, vitamins, and amino acids that provides the necessary nutritional requirements for cultured plant cells and tissues to grow. It contains macro and micro nutrients, vitamins, amino acids or other nitrogen supplements, sugars or other organic supplements, solidifying agents, and plant growth regulators.
Mutation & its detection
1. The document discusses mutation and its detection. It defines mutation as heritable changes in the genome excluding those from other organisms.
2. It describes different types of mutations such as spontaneous versus induced, forward versus reverse, nuclear versus cytoplasmic, and more.
3. Methods of detecting mutations in prokaryotes and eukaryotes are described. For prokaryotes, techniques like replica plating and the Ames test are used. For eukaryotes, each individual must be examined for mutant phenotypes.
Somatic hybridization faizy
This document provides an introduction to somatic hybridization, which is the fusion of somatic protoplasts from two different plant species or varieties to create hybrid plants. It discusses the history and development of the technique, including early work isolating protoplasts. The key steps in somatic hybridization are described: isolating protoplasts using either mechanical or enzymatic methods, fusing the protoplasts spontaneously or through induction methods, identifying and selecting hybrid cells, culturing the hybrid cells, and regenerating hybrid plants. Advantages include creating novel hybrids and transferring desirable traits between species, while limitations include low regeneration rates and viability of fused products.
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Sexual reproduction in bacteria involves the transfer of genetic material between bacterial cells through three main mechanisms: transformation, transduction, and conjugation. Transformation involves the uptake of naked DNA by competent bacterial cells. Transduction occurs when bacterial DNA is transferred by bacteriophages. Conjugation is the transfer of DNA through direct cell-to-cell contact via conjugation pili. Gram staining is a technique used to differentiate between Gram-positive and Gram-negative bacteria based on differences in cell wall structure.
Protein targetting ppt (1)
This document summarizes protein targeting mechanisms in cells. It discusses how signal sequences direct proteins to different organelles like the endoplasmic reticulum, mitochondria, chloroplasts and nucleus. The signal sequence is cleaved after the protein reaches its destination. Glycosylation in the ER plays a key role in targeting lysosomal enzymes. Mitochondrial and chloroplast proteins use a different targeting mechanism after full synthesis. Receptor-mediated endocytosis imports some extracellular proteins by binding to receptors and forming clathrin-coated vesicles.
Anther culture
Anther culture is a technique where anthers are excised from flower buds and cultured on nutrient media. This can produce haploid plantlets through either organogenesis or embryogenesis. The first report of haploid tissue from anther culture was in 1966 in Datura pollen grains. Haploids are useful for plant breeding as they contain only one allele per gene, revealing recessive traits, eliminating lethal genes, and allowing for efficient production of homozygous plants. The protocol involves sterilizing tobacco flower buds, removing anthers, and culturing them on nutrient media. After 3-4 weeks, haploid plantlets emerge from the cultured anthers. Haploids have applications in basic research, mutation studies
totipotency
Totipotency refers to a plant cell's genetic potential to regenerate an entire plant. In plant tissue culture, cells taken from plant parts can form an unorganized mass of cells called callus tissue when placed in a sterile nutrient medium. The cells within callus tissue are totipotent, meaning they have the ability to regenerate into a whole plant under the right conditions and hormone manipulations. Totipotency is important for plant science applications like vegetative propagation of crops, genetic modification of plants, and preservation of endangered plant species through long-term storage of totipotent cells.
Anther culture
Anther culture is an in vitro technique used to produce haploid plants from male gamete cells. Haploid cells contain only one set of chromosomes. The first report of anther culture was in the 1970s, while the first natural occurrence of haploids was observed in 1922 in Datura plants. Anthers containing microspores are cultured on nutrient media supplemented with hormones and sugars. Depending on the species, haploid plants or callus can regenerate from the cultured anthers within 3-8 weeks. Haploid plants are useful for breeding programs as they can be doubled to generate fertile, homozygous plants in one generation.
PROTOPLAST ISOLATION & FUSION
This document discusses protoplast isolation and fusion. It defines a protoplast as a plant, bacterial, or fungal cell that has had its cell wall removed, leaving the plasma membrane intact. The document outlines the history of protoplast isolation, from the first isolation in 1892 to the use of commercial enzyme mixtures in the 1960s. It also describes how protoplasts can be fused using polyethylene glycol or other fusogens to create somatic hybrids for plant breeding purposes.
Special type of chromosomes
This document discusses two special types of chromosomes: polytene chromosomes and lampbrush chromosomes. Polytene chromosomes are found in salivary glands and are formed through repeated DNA replication without cell division, resulting in fused sister chromatids. Lampbrush chromosomes are found in growing oocytes of birds and some animals, and appear during meiosis with large chromatin loops and chromomeres that give them a lampbrush-like appearance used for active gene transcription. Both special types of chromosomes are much larger than normal chromosomes and are involved in high levels of gene expression or RNA transcription.
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A presentation covering the process of protoplast culture including protoplast isolation, protoplast fusion, culture of protoplast, its application, factors affecting protoplast culture and the future of protoplasts.
Protoplast fusion
Protoplast fusion involves removing the cell walls of plant cells through enzymatic or mechanical means to create naked protoplasts. These protoplasts can then be fused using chemicals, electricity, or other methods. This allows the cytoplasms and sometimes nuclei of different plant cells to merge, creating hybrid cells. Successful fusion can generate hybrid plants through regeneration of cell walls and tissues. Protoplast fusion overcomes sexual incompatibility and is used to introduce traits like disease resistance between species. It remains a technically challenging process with limitations like genetic instability and uncertain expression of transferred traits.
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Suspension Culture and Single Cell Cultures, Culturing methods, maintenance and application
Generally, suspension culture is a one stop technology to produce secondary metabolites on a large scale in-vitro, irrespective of the climatic condition or nutrient availability (as required in field plants).
In this presentation, we will see the importance of suspension culture, culturing methods and it's application (mostly with respect to plants) and also focus on what exactly is a single cell culture.
Development in arabidopsis
Introduction And Classification
Anatomy Of Flower
Life Cycle Of Arabidopsis
Early Flower Development
Embryogenesis-
A. Formation Of Microspores
B. Formation Of Megaspores
Embryonic Development Starts By Establishing A Root-shoot Axis And Then Halts Inside The Seed
Arabidopsis Genome Is Rich In Developmental Control Genes.
Control Of Carpel & Fruit Development
Arabidopsis Thaliana A Model Plant
Conclusion
References
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Lecture 20 superovulation and embryo transfer in cattle
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Elektroporasyon cihazi
- 1. 1 T.C. ERCİYES ÜNİVERSİTESİ MÜHENDİSLİK FAKÜLTESİ BİYOMEDİKAL MÜHENDİSLİĞİ Samet ÇIKLAÇANDIR 1
- 2. 2 Elektroporasyon nedir? Referans cihazımız ve aksesuarları Elektroporasyonun kullanıldığı alanlar Proje aşamalarımız
- 3. 3 Elektroporasyon gen aktarım yöntemlerinden biridir. Bu işlemde hücrelere veya dokulara kısa süreli ve çok kuvvetli elektrik voltajı uygulanır. Buna bağlı olarak hücre zarında küçük çapta geçici porlar (delikler) oluşur. Bu deliklerden de DNA’lar hücreye aktarılır. Şekil: Aşamalar Sırasıyla
- 4. 4 REFERANS ALDIĞIMIZ CİHAZ VE AKSESUARLARI Footswitch 45-0211 GenetrodesTweezertrodes
- 5. 5 NERELERDE KULLANILIR? Cilt hücrelerinin yenilenerek genç kalmasında Genetik yapı değişikliğinde Bazı ilaçların doğrudan hücre içine aktarılmasında Daha yüksek voltaj uygularsak kanser tedavisinde Meyvelerden meyve suyu yapımını %10 arttırılması
- 6. 6
- 7. 7 Şekil: Burada da daha düşük voltajla tümörlü hücrenin içine ilaç sokularak hücrenin ölmesine yol açılmış.
- 8. 8 PROJE AŞAMALARIMIZ Elektroporasyon yöntemi ve elektroporatör üzerine teorik bir araştırma yaptık. Devre için gerekli elektronik devre elemanlarını belirledik ve çizim programı üzerinde devremizi çizdik. Programdan istediğimiz çıktıyı aldık. Voltaj düşürücü transformatör Artırım tip N kanallı MOSFET
- 9. 9 Darbe sayısı, darbe süresi ve duty cycle ayarı için PIC programı yazarak PIC16F877A’ya yükledik. Bu parametreleri gösteren LCD’yi de programladık. PIC16F877A LCD GÖSTERGE
- 10. 10 Şekil: İlk Yaptığımız Katlama İşlemi
- 11. 11 Şekil: PIC C Compiler ile mikrodenetleyici programı yazdık. Bu, uygulanacak kısa süreli sinyalin kontrolü için yapılır. Şekil: Pic İle Anahtarlama İşlemi ve Darlington Bağlantısı
- 12. 12 Şekil: Pic’i Beslemek İçin Şebeke Geriliminin Trafo ile Düşürüp Doğrultulması Şekil: Pic ile Periyot (Pulse Uzunluğu) Pulse Sayısı Duty Cycle Oranı Ayarları Yapılırken
- 13. 13 Şekil: Proteus’da Devrenin Son Hali
- 14. 14 Şekil: 2000 ms Uzunluklu 3 Adet %10 Duty Cycle Uygulanıyor Şekil: 410 ms Uzunluklu 4 Adet %50 Duty Cycle Uygulanıyor
- 15. 15 Bilgisayar üzerinde tasarladığımız devreyi protoboarda aktardık. Şekil: Protoboarda aktarılmış devre
- 16. 16 Şekil: Anahtar, mikrodenetleyici ve LCD’nin boarda aktarılmış hali Şekil: Güç mosfeti ile anahtarlama devresi
- 18. 18 Şekil: Devrenin elemanlarla gerçekleştirilmesi Şekil: Katlama sonucu 1119 volt multimetrede görüldü.
- 19. 19 Aktardığımız devrenin çıktılarını osiloskop üzerinde aldık, doğru olduğunu teyit ettik. Şekil: Devremizden osiloskop üzerinden çıktı alınması
- 20. 20 • Darbe süresi : 20 milisaniye • Darbe adedi : 4 • Darbe genişliği : %50 Şekil: Yukarıda belirtilen darbe özelliklerine göre çıkışın osiloskopta görüntüsü
- 21. 21 • Darbe süresi : 20 milisaniye • Darbe adedi : 5 • Darbe genişliği : %80 Şekil: Yukarıda belirtilen darbe özelliklerine göre çıkışın osiloskopta görüntüsü
- 22. 22 • Darbe süresi : 32.2 milisaniye • Darbe adedi : 3 • Darbe genişliği : %90 Şekil: Yukarıda belirtilen darbe özelliklerine göre çıkışın osiloskopta görüntüsü
- 23. 23 • Darbe süresi : 45 milisaniye • Darbe adedi : 7 • Darbe genişliği : %30 Şekil: Yukarıda belirtilen darbe özelliklerine göre çıkışın osiloskopta görüntüsü