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What treatment do cells require to be competent?
this depends on whether you are trying to make chemically competant cells or electrically competant cells. the process of making a cell "competant" means that they are capable of accepting 'donor' DNA
What is the definition of competent cells and how it can help to make a transformation more efficient during the cloning of desired gene?
Generally most of the bacteria have the ability to take up DNA which is present freely in the environment. This is called Competence. It is genetically acquired. But some cells do not exhibit natural competence and they are treated with a suitable chemical like CaCl2 to make their cell wall relatively permeable to DNA. We can make any cell competent in vitro and introduce our desired gene which is taken up by the competent cell. To test the efficiency of our treatment, we may introduce a marker gene like gene for antibiotic resistance such as Tetracyclin onto the desired gene and grow the treated cells on Tetracyclin medium so that only the cells that have taken up the DNA survive.
What did Fred Griffith discover when he injected mice with a mixture of dead pathogenic S cells and living harmless r cells of Streptococcus pneumonia?
Fred Griffith discovered that the harmless R strain of Streptococcus pneumoniae could be transformed into the deadly S strain when mixed with heat-killed S strain bacteria. This experiment provided evidence for the concept of bacterial transformation, where genetic material can be transferred between different strains of bacteria.
Why gene gun use in rice transformation?
It's a tool to insert foregin gene into target cells and can be used for any suitable crop or cells for transformation purpose. Now a days, agrobacterium mediated transformation techniques are becoming popular.
What is roll of calcium chloride in making competent cells in transformation?
The addition of calcium chloride to a cell suspension promotes the binding of plasmid DNA to Lipopolysaccharide. Positively charged calcium ions attract both the negatively charged DNA backbone and the negatively charged groups in the Lipopolysaccharide inner core. The plasmid DNA can then pass into the cell upon heat shock, where cells are cooled to a low temperature (+4 degrees Celsius) and then heated to a high temperature (+42 degrees Celsius) for a short time.
What is meant by term competent when applied to bacteria being prepared for transformation?
Being competent refers to the ability of bacteria to uptake foreign DNA. In the context of transformation, bacteria are made competent through the use of special techniques that temporarily disrupt their cell walls, allowing foreign DNA to enter the cells. This process is essential for the successful transfer of new genetic material into bacterial cells.
What is the definition of competent cells?
cells which readily accept a foreign DNA through a process called transformation
Can cells be transformed when they are in a competent state?
Yes, cells can be transformed when they are in a competent state. Competent cells have an increased ability to take up foreign DNA, making them more likely to undergo transformation when exposed to external DNA molecules. This process is commonly used in laboratories for genetic engineering and cloning.
What are the reason for the low level of transformation during the experiment?
Some possible reasons for low transformation efficiency during an experiment could include low competency of the host cells, poor quality of plasmid DNA, improper handling of cells during transformation, and suboptimal growth conditions post-transformation. It's important to troubleshoot each step of the protocol to identify the specific cause of low transformation efficiency.
How molecular mechanism of bacterial transformation is carried out?
Basically, transformation refers to the uptake and incorporation of naked genetic material by a competent cell. Most bacterial cells are not naturally competent(able to take up naked genetic material). The kinds that are use a type 4 pilli to get the genetic material into the cell. This process is very rare in nature, since it requires naked genetic material to be present, from a lysed cell, as well as a competent cell to take up the genetic material. When the DNA/RNA is taken up, it is often degraded by restriction enzymes. If it is not degraded it can exist as extra chromosomal DNA, much like a plasmid, or it can be incorporated into the cells chromosome. Scientists use heat, pH, or osmotic shock in lab to make cells artificially competent.
Why do you keep competent cells in ice?
Competent cells are stored in ice to maintain their viability and stability. Storing them at low temperatures helps prevent degradation of their membrane and genetic material, ensuring that they remain suitable for genetic transformation experiments.
What is meant by competent cells?
Competent cells are bacterial cells that have been treated to make them capable of taking up foreign DNA, such as plasmids. These cells can be used for various genetic engineering techniques, such as transformation.
What treatment do cells require to be competent?
this depends on whether you are trying to make chemically competant cells or electrically competant cells. the process of making a cell "competant" means that they are capable of accepting 'donor' DNA
What is LB plate in making competent cells?
LB plate is a commonly used nutrient agar plate that contains Luria-Bertani (LB) broth and agar, which provides essential nutrients for bacterial growth. LB plates are used in the production of competent cells by providing a suitable environment for bacterial culture growth and transformation efficiency.
What is The transfer of DNA into cells?
Transformation
What is the definition of competent cells and how it can help to make a transformation more efficient during the cloning of desired gene?
Generally most of the bacteria have the ability to take up DNA which is present freely in the environment. This is called Competence. It is genetically acquired. But some cells do not exhibit natural competence and they are treated with a suitable chemical like CaCl2 to make their cell wall relatively permeable to DNA. We can make any cell competent in vitro and introduce our desired gene which is taken up by the competent cell. To test the efficiency of our treatment, we may introduce a marker gene like gene for antibiotic resistance such as Tetracyclin onto the desired gene and grow the treated cells on Tetracyclin medium so that only the cells that have taken up the DNA survive.
What do control setup and experimental setup have in common?
Everything except what you're measuring. EXAMPLE: In a biological experiment using specially treated cells the experiment is run with the cells; this is the experiment. Then the experiment is run again with the same cells WITHOUT the special treatment; this is the control.
