Janith Piumal

Janith Piumal

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Blotting Techniques

Blotting Techniques mainly have four main steps Electrophoretic separation of protein/nucleic acid fragments in the sample Transfer to and immobilization on a solid support Binding of probe to the target molecule on membrane Visualization of bound probe Southern blotting –…

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Electrophoresis of Nucleic Acids.

Electrophoresis − the migration of a charged particle under the of an electric field. Migration rate/separation depends on: size, shape, charge The charge per unit length in any given fragment of DNA is the same since mobility of DNA molecules…

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DNA/RNA QUANTIFICATION

DNA and RNA quantitation is performed using various methods. Absorbance/spectrophotometric methods Gel electrophoresis – Agarose gel electrophoresis Fluorometric methods Absorbance method (UV spectrophotometer, NanoDrop) At 260 nm nucleic acids absorbs light most strongly At 280 nm proteins are absorbs light…

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Plasmid DNA extraction.

Plasmid DNA may appear in several conformations. Covalently closed circular DNA (cccDNA) – both strands of the plasmids DNA are intact. (Supercoiled DNA, Relaxed circular DNA) Open circular DNA – One nucleotide stand is broken Super coiled DNA is move…

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RNA Extraction.

RNases enzyme is one of the main problem of the RNA extraction. that degraded the RNA because RNases is very stable, found every everywhere, active enzyme that do not required any co-factors and maintain activity even during the autoclaving. Avoiding…

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DNA extraction

The key steps of DNA extraction is give below. Sample disruption Clearing cell debris Purification Concentration DNA Sample disruption. rapid and complete disruption of cells in sample releasing DNA into lysate. there are several methods to disruption of the cell…

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Plant Improvement.

The goal of plant improvement is to create superior plants from existing ones (higher yield, improved quality, resistance to abiotic and biotic stress). Ability of a particular trait to be manipulated is determined by environment and the genotype. Heritability is…

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Plant Propagation.

Plant propagation is the creation of new plants from existing ones. There are two main types of plant propagation: asexual (vegetative propagation), sexual (seed propagation). Genetic control in propagation – in vegetative propagation plants are genetically similar to their parents.…

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Germplasm Conservation.

Germplasm is the living resource for genetic information in a plant. pass from one generation to another. Germplasm conservation is the preservation of these genetic information. Genetic erosion – Decline in the genetic variation in cultivation or natural population. Causes…

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Breeding methods for self pollinating plants.

The following are the main breeding methods for self pollinating plants: Mass selection Pure – line selection Hybridization Marker assisted selection synthetic populations Mass selection Mass selection is the process of selection of the crops breeding based on their overall…