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In molecular biology, a hybridization probe is a fragment of DNA or RNA of variable length (usually 100-1000 bases long), which is used in DNA or RNA samples to detect the presence of nucleotide sequences (the DNA target) that are complementary to the sequence in the probe. The probe thereby hybridizes to single-stranded nucleic acid (DNA or RNA) whose base sequence allows probe-target base pairing due to complementarity between the probe and target. The labeled probe is first denatured (by heating or under alkaline conditions such as exposure to sodium hydroxide) into single stranded DNA (ssDNA) and then hybridized to the target ssDNA (Southern blotting) or RNA (northern blotting) immobilized on a membrane or in situ.
To detect hybridization of the probe to its target sequence, the probe is tagged (or labelled) with a molecular marker of either radioactive or (more recently) fluorescent molecules; commonly used markers are 32P (a radioactive isotope of phosphorus incorporated into the phosphodiester bond in the probe DNA) or Digoxigenin, which is non-radioactive antibodybased marker. DNA sequences or RNA transcripts that have moderate to high sequence similarity to the probe are then detected by visualizing the hybridized probe via autoradiography or other imaging techniques. Normally, either X-ray pictures are taken of the filter, or the filter is placed under UV light. Detection of sequences with moderate or high similarity depends on how stringent the hybridization conditions were applied — high stringency, such as high hybridization temperature and low salt in hybridization buffers, permits only hybridization between nucleic acid sequences that are highly similar, whereas low stringency, such as lower temperature and high salt, allows hybridization when the sequences are less similar. Hybridization probes used in DNA microarrays refer to DNA covalently attached to an inert surface, such as coated glass slides or gene chips, and to which a mobile cDNA target is hybridized.
Depending on the method the probe may be synthesized using phosphoramidite method or generated and labeled by PCR amplification or cloning (older methods). In order to increase the in vivo stability of the probe RNA is not used, instead RNA analogues may be used, in particular morpholino. Molecular DNA- or RNA-based probes are now routinely used in screening gene libraries, detecting nucleotide sequences with blotting methods, and in other gene technologies such as microarrays. When people think of the word 'cloning' they are often hit with frightening images of duplicate human beings being created in somewhat of a mad scientist style experiment. In fact, many members of the public were outraged when Dolly the sheep resulted from a cloning experiment in Scotland. Therapeutic cloning, however, is entirely different and does
which had its nucleus previously removed. Related on Explore Stem Cells. The nucleus holds the genetic material for a human or laboratory animal. The cells in the inner cell mass are isolated and then utilised to create embryonic stem cell lines. Overview of Stem Cell Therapy Stem Cell Research Around the World Ovarian Function and Stem Cells Stem Cells to Treat Blindness Disappointments in Stem Cell Therapies MS and Stem Cells Benefits of Therapeutic Cloning A major benefit of therapeutic cloning is that the cells removed are pluripotent. imparting structure and function as needed. The nucleus that is extracted from the somatic cell in the patient is then inserted into the egg. In a very basic sense. In practical human applications. In this procedure. called the inner cell mass that is rich in stem cells. it's a procedure of substitution. In therapeutic cloning. or instructions. How is Therapeutic Cloning Performed? Therapeutic cloning is another phrase for a procedure known as somatic cell nuclear transfer (SCNT). Pluripotent cells can give rise to all cells in the body with the exception of the embryo. which is any body cell other than an egg or sperm..not involve the creation of a perfectly copied human being. Scientists then take a somatic cell. It is stimulated to divide and shortly thereafter forms a cluster of cells known as a blastocyst. The egg now contains the patient's genetic material. a researcher extracts the nucleus from an egg. the somatic cell would be taken from a patient who requires a stem cell transplant to treat a health condition or disease. This blastocyst has both an outer and inner layer of cells and it is the inner layer.. and also extract the nucleus from this cell. This means that . It is reproductive cloning that results in a copy of a specific human being. no sperm fertilisation is involved nor is there implantation into the uterus to create a child. which are infused into the patient where they are ideally integrated into the tissues.
support therapeutic cloning for treating disease. The majority of scientists are adamantly opposed to reproductive cloning and instead. The ultimate consequence would be a rejected stem cell transplant. Another distinct advantage to this type of therapy is that the risk of immunological rejection is alleviated because the patient's own genetic material is used. Therapeutic cloning does result in the destruction of an embryo after stem cells are extracted and this destruction has stirred controversy over the morality of the procedure. This understanding can hopefully lead to new treatments or cures for some of the common diseases affecting people today. Problems with Therapeutic Cloning One problem with therapeutic cloning is that many attempts are often required to create a viable egg.pluripotent cells can potentially treat diseases in any body organ or tissue by replacing damaged and dysfunctional cells. the procedure would allow for scientists to create stem cell therapies that are patient specific and perfectly matched for the patient's medical condition. Therapeutic cloning is also important to enhancing our understanding of stem cells and how they and other cells develop. This means that therapeutic cloning has the potential to dramatically reduce the wait times for organ transplants as well as the immunological concerns associated with organ transplant therapy. This is one of the major challenges of organ transplants. In addition. the patient's body would be more likely to recognise the foreign proteins and then wage an attack on the transplanted cells. To this date. a scientist may attempt to move beyond therapeutic cloning to creation of a human being. but also for most of the ethical scientific field. The stability of the egg with the infused somatic nucleus is poor and it can require hundreds of attempts before success is attained. Because reproductive cloning does utilise SCNT as the primary step. alongside the fact that there is a huge shortage of available organs for those who require the procedure. no human being has been successfully cloned but the possibility of this occurring is a frightening one not only for the general public and policy makers. If a cell line were created with cells from another individual. Some argue that the pros outweigh the cons with regards to treating disease whilst others have likened the destruction to an abortion. Still others state that this doesn't change the fact the embryo could potentially be a human being and so destruction of the embryo is no different than destruction of a human life. there is also still fear that given our knowledge base to perform reproductive cloning. With policies and careful monitoring in place to ensure that therapeutic cloning is used .
MA. Adult DNA cloning cannot ethically be used to produce a human clone. The initial steps of the procedure were tried using human DNA in 1998DEC." This causes a great deal of confusion among the public. or perhaps one day cure. Many transfer their disgust at the concept of creating cloned babies to therapeutic cloning whose goal is to create an organ to heal people.a. many diseases.k. a pro-life senator has suggested the term "somatic cell nuclear transfer" to replace therapeutic cloning. cell nuclear replacement): This involves removing the DNA from an embryo and replacing it with the DNA from a cell removed from an individual.k.responsibly. or quantity of skin. Three very different procedures have been referred to as "cloning. loning is the production of one or more individual plants or animals (whole or in part) that are genetically identical to an original plant or animal. The end result would not be a human being.a. a biotech company in Worcester. Orrin Hatch (R-UT). because experiments on animals have sometimes produced defective specimens. or piece of nerve tissue. This method has been used to clone a sheep. it would be a replacement organ. both terms contain the word "cloning. although adult DNA cloning and therapeutic cloning are quite different procedures leading to very different goals. It's stem cells would then be extracted and encouraged to grow into a piece of human tissue or a complete human organ for transplant. The first successful therapeutic cloning was accomplished in 2001-NOV by Advanced Cell Technology. Therapeutic cloning: (a. the embryo would be implanted in a woman's womb and be allowed to develop in to a new human whose DNA is identical to that of the original individual. Unfortunately. The resultant embryo would be allowed to grow for perhaps 14 days." Two are: Adult DNA cloning (a. 3 How is therapeutic cloning done? . Then. Somatic cell nuclear transfer or research cloning): This starts with the same procedure as is used in adult DNA cloning. we can all benefit from the potential of this procedure to eventually treat.
and about a dozen empty flower pots. However. What is involved in somatic cell nuclear transfer is to: Take a woman's ovum. this results in its death. Stem cells are a unique form of human cell that can theoretically develop into many organs or body parts the body. and inserting it into the ovum.I once had a Swedish ivy plant. In a small percentage of cases. Each cell is. In a few weeks. I cut off many leaves from the original plant. Giving the resulting ovum an electrical shock to start up its embryo making operation. Actually. 4 An important factor to remember is that: . the pre-embryo is not implanted in a woman's womb in order to try to produce a pregnancy. The stem cells would be encouraged to grow into whatever tissue or organ is needed to treat the patient. A woman's ovum also contains her DNA. and surrounded the pots with clear plastic. So too. a pre-embryo will be formed. DNA testing on an human often starts by scraping some cells from the inside of a person's mouth. a form of human life for the simple reason that it contains human DNA. it contains the DNA of the person. The pre-embryo is allowed to develop and produce many stem cells. placed each of them in a pot filled with damp soil. Remove the DNA from a cell taken from a human. and thus contains all of the information required to produce a duplicate or cloned person. No matter how a cell is obtained. I had a dozen new plants. all the information needed to create a new human being is contained in each cell of an existing human being. This shows that all of the material and information needed to create a Swedish ivy plant is contained in a single leaf. The tissue or organ would be transplanted into the patient. Living cells can be scraped off of a person's skin. This converts it to a form of human life into what is basically a factory for creating a pre-embryo. in fact. So far. the procedure is identical to that used in adult DNA cloning. it is contained in a single cell of a single leaf. and remove its DNA. Stem cells are removed from the pre-embryo.
it contains human DNA. it contains human DNA. etc) Therapeutic cloning has not yet been accomplished in the laboratory or clinic." 4 If scientists are successful. whether it comes from a skin scraping or is extracted from the inside of a person's mouth. skin. liver. The pre-embryo that is produced is a form of human life." This has been done for most of the 220 cell types in the human body. then perfectly matched. pancreas. to the pre-embryo. The transplanted tissue must develop normally and must not represent significant "risks to the patient. it contains human DNA. injuries. or disorders." This has already been accomplished They have to be encouraged to "turn into specific cell types. Future experiments may not succeed. There were four main hurdles to overcome: Stem cells have to be "successfully isolated and grown in the laboratory. replacement organs could become freely available to sick and dying people. The original ovum is a form of human life. there is a continuity of human life from a surplus cell which a human produces by the millions each day.The original seed cell is a form of human life. it would probably take many years of research before the first useful results will be obtained. Theoretically. these stem cells can be used to develop into replacement organs (heart. What are its possible benefits? If therapeutic cloning using embryos is successful. They have to be proven usable in treating patients with diseases. That would save countless . So.
For transplants involving kidney (or theoretically any other organ that is duplicated in the body).numbers of lives. Down Syndrome. inconvenience. pediatrics and reproductive medicine--recently sent a letter to members of Congress urging them to support federal funding for. and increase the quality of life of countless others. and potentially shortened life span in order to donate the organ. cystic fibrosis. when compared to regular organ transplant donated by a second person: There would be presumably be no danger of rejection of the transplant because the organ's DNA would match the patient's DNA exactly. The potential exists to cure. ophthalmology.. AIDS. "More than 50 disease advocates and scientific societies.. a brand-new organ would be grown specifically for them. The patient would not have to wait until an unrelated donor dies to obtain a transplant. "Further advances in understanding of how organs regenerate would increase the range of possible treatments that could be considered." 4 There would probably also be side benefits resulting from the research. Parkinson's disease. lymphoma. another individual would not have to experience pain. certain diseases and disorders that cannot be effectively handled today. aging. A new organ could be grown for them as needed. The procedure would save lives which would otherwise be lost waiting for a transplant that did not come in time. or at least treat." 4 In the United States during 1998. Three possible examples of therapeutic cloning "might include the use of insulin-secreting cells for diabetes. infertility and cancer--as well as professional groups that focus on such issues as cell biology. blindness. representing such concerns as diabetes. The patient would not have to make-do with a replacement organ that is old and may have reduced functionality." 5 This procedure would have a number of advantages. microbiology. nerve cells in stroke or Parkinson’s disease. What are the problems of therapeutic cloning: . or liver cells to repair a damaged organ. glaucoma. cardiology.[stem cell] research. stroke.
Some believe that somatic cell nuclear transfer is sufficiently similar to normal conception with an egg and spermatozoa that a human person also comes into existence during therapeutic cloning. In other cases. Stability of stem cells: As of 2003-MAR. 15 million eggs would be required to completely wipe out diabetes. It is obvious that therapeutic cloning will not be feasible until these deficiencies have been overcome. very few cures could be made for economic reasons. Stem cells have sometimes mutated. Extracting eggs from women is "painful. a Fundamentalist Christian group. this would still require over a half million . diabetes. they have produced tumors. it would take 1. have shown great promise. Until the production of stem cells becomes more efficient. and perhaps Canada. at least with experiments on animals. Unfortunately. the embryo..Before therapeutic cloning can become generally available to cure heart disease. Even if and when techniques are found to reliably produce one custom stem cell line for each egg harvested from a woman. a number of hurdles have to be overcome: Developing cures: Methods have to be developed that will cure or treat diseases with embryonic stem cells. to cure another person of paralysis. which has been underway for many years. this is murder. like China. 8 We have been unable to determine what form this injury might take. the UK. paralysis. therapeutic cloning is still in its early stages of development. chief executive of Geron Corporation -. or heart disease.5 billion eggs to cure the 15 million Americans who have diabetes. This looks promising. They feel that murdering one person. etc. Assuming two dozen eggs per woman. and thus been rejected by the recipient's body. Thomas Okarma." 7 Focus on the Family. costly and unreliable. or diabetes. can never be justified. Research using therapeutic cloning is a new field. but it has already shown that stem cells from embryos have much greater flexibility than adult stem cells. cited an unknown expert who has said that the process of harvesting eggs would seriously injure about one percent of all female donors. Is an embryo a human person? Pro-life supporters generally believe that a human person comes into existence at conception. More details. 6 This means that if a cure for diabetes involving therapeutic cloning is found. Research with adult stem cells. Where would the eggs come from? At the present stage of somatic cell nuclear transfer. The process of extracting stem cells involves killing the embryo. Therapeutic cloning research may well be limited to those countries. adult cells are limited in their application. etc.estimated that takes "100 eggs if you're lucky" to produce a useable stem cell line. where pro-life supporters are relatively few in number.a leading stem-cell research establishment -. To many pro-lifers.
Weiner. The Rhesus factor is an antigen. If the woman is Rh-negative and has been sensitized. or delivery. people that don't have the antigen in their blood. So a person who has a blood type of A and has the Rhesus factor is said to have A-positive. Rapid.COM If the mother is Rh-negative and the child is Rh-positive. Subsequent pregnancies will likely require separate Rhlg injections. "the embryo will lose all traces of its rabbit origin. So.bizbuysell. IgG. it is generally recommended that the newborn be tested for his or her Rhesus blood type. This should prevent sensitization for the rest of the pregnancy. Specifically. that is. That is. that is.com Bath Kneeler for Mom Bathing Baby Gift for New Mom.org Looking for a Educational Business? View Businesses in Michigan. are Rh-negative. the woman's body may naturally produce antibodies which attack the baby's blood. Cute Exercise Pad WWW. People with the Rhesus factor. or more specifically a protein. IgE. are Rh-positive. also known as the Rh factor. Close monitoring of the baby should be conducted to ensure that Rh disease is not developing. that is. If the woman has become sensitized. Each blood type is further labeled as positive or negative. A person's Rh type is generally significant only with respect to pregnancies. the mother is often given another Rhlg shot shortly after birth to prevent her from becoming sensitized." The Rhesus factor. a Rh-positive child born to an Rh-negative woman runs the risk of developing Rh disease. the father must have been Rh-positive. brain or heart damage. Preventative measures to protect against the Rhesus factor disease and their devastating effects are available. including miscarriages.women willing to donate eggs and run the risk of some type of injury. gets its name from experiments conducted in 1937 by scientists Karl Landsteiner and Alexander S. Rh-positive women do not. A Rh-positive man has a 50% chance of passing on his Rh-positive blood type to the child All forms: IgA. Therefore. . and AB. Potential health problems include jaundice. blood. O. anemia. Additionally. Rhlg injections only last for a given pregnancy. These revolutionary case studies involved rabbits which. Sensitization may not only occur through normal pregnancies.bioatla. B.right4eu. but at any time a woman and her child's blood mix. The Rhlg shot seeks to destroy any antigens present in the bloodstream before the mother is able to create antibodies.eclt. those antibodies will be present in her system for the rest of her life. Blood transfusions to replace the diseased blood with healthy blood may be given during or after delivery depending on the circumstances. If a woman is Rh-negative. The Rhesus factor is less likely to affect the first-born child because the woman's system will have had less time and is therefore less likely to produce the antibodies to fight off the antigens in the child's Rh-positive blood. people with the antigen present in their blood. During the process. People without the Rhesus factor. her immune system responded by producing antibodies. The result of this incompatibility will not affect the health of the mother but it can affect the child's health. the Rhlg injection will not help. IgM Proven. labor. Therapeutic cloning will probably only become generally useful when a method is found to use non-human eggs as source material. Women should be tested early in their first pregnancies to determine whether they are Rh-negative and whether they are sensitized. and in severe cases Rh disease can be fatal. causing the baby's red blood cells to break down. or A+. that exists on the surface of red blood cells. High Quality www. which is a reference to the Rhesus factor of the blood. and blood transfusions. for a Rh-negative woman to have a Rh-positive child. ectopic pregnancies. disparate Rhesus factor types between a woman and her child can increase the potential for Rh disease in each subsequent pregnancy. If the child is positive. produced an antigen present in the red blood cells of many humans. IgD. she will likely be given an injection of a blood product known as Rh immunoglobulin (Rhlg) at about seven months into the pregnancy. the woman's immune system may respond by producing antibodies to fight off the child's antigens which are foreign to the woman's system. More than 85% of people are Rhpositive. Books and Test Kits www. Only Rh-negative women risk having children with the Rhesus factor disease.CuteAndComfyCushionKnee. when injected with the Rhesus monkey's red blood cells. There are four general categories of blood: A. and the child's blood enters the woman's bloodstream during the pregnancy.com Blood Type Diet Vitamins. and has not yet been sensitized. Research is underway to use rabbit eggs.com A unique alliance to address child labour in tobacco growing www. www.
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