Diagnosis of Beta-Thalassaemia by Dna Amplification in Single Blastomeres From Mouse Preimplantation Embryos

Holding, Cathy; Monk, Marilyn

doi:10.1016/s0140-6736(89)90655-7

Cited by 87 publications

(46 citation statements)

References 7 publications

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“…Finally, lanes 7-9 show the result of carrying out the same procedure used in lanes 4-6 except that the second round of amplification on the aliquot involved replacement of one of the original primers with an internal primer, a procedure we call heminesting. Heminesting is a modification of the "nesting" method (5,10), in which a second round of amplification uses two new amplification primers specific for the same target, both of which are internal to the first primer pair. While the introduction of the Taq DNA polymerase eliminates the need for nesting in most PCR applications, we find heminesting extremely valuable for the analysis of single cells.…”

Section: Resultsmentioning

confidence: 99%

“…For each locus to be tested, 10 .l of the lysed sample (equivalent to 0.6 ,l of semen and containing approximately 5 x 104 sperm) was directly subjected to the ADPL procedure without the initial amplification step. As discussed above, parallel experiments were carried out to confirm the ADPL results by using an ASO hybridization assay.…”

Section: Resultsmentioning

confidence: 99%

“…By using a "full" nesting technique (5) the ,8-globin gene has been electrophoretically detected with little background in single mouse blastomeres; no signal was detected from eggs of mice with a globin gene deletion (10). By using allele-specific primers of different lengths in a heminesting protocol we not only eliminate background PCR products but are capable of analyzing the allelic state at three independent loci simultaneously in a single cell.…”

Section: Resultsmentioning

confidence: 99%

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Direct electrophoretic detection of the allelic state of single DNA molecules in human sperm by using the polymerase chain reaction

1990

Trends in Genetics

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We have developed a procedure that allows the detection of polymerase chain reaction (PCR) products derived from a single target DNA molecule in a human sperm without using radioactive probes. With this method, three genetic loci present in a single sperm can be amplified simultaneously. The amplification procedure is specific as well as efficient and permits detection of the PCR product by ethidium bromide staining after polyacrylamide gel electrophoresis. When allele-specific PCR primers that differ in length are used, the size of the PCR products of different alleles also vary in length, allowing the allelic state at each locus to be determined electrophoretically. Studies on individual sperm by using this procedure should facilitate the measurement of genetic recombination in humans over small physical distances. The ability to directly analyze the allelic state of PCR products from one cell rapidly and simply will also be useful for the prenatal diagnosis of genetic disease, especially in the analysis of single blastomeres taken from in vitro fertilized eggs prior to implantation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Direct electrophoretic detection of the allelic state of single DNA molecules in human sperm by using the polymerase chain reaction

1990

Trends in Genetics

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“…It is noteworthy to mention that Dr. Ken White's doctoral research was presented at the 1982 International Embryo Transfer Society meeting and was the recipient of the first inaugural Student research award [8]. Ultimately, the emphasis on genetic determination led to the development of preimplantation diagnosis (PGD) in the mouse model [9,10]. In conjunction with the specific polymerase chain reaction (PCR) testing for X and Y chromosomes [11][12][13], simultaneous efforts to improve the efficacy of single cell biopsying [14] facilitated human PGD development [15,16].…”

Section: Genetic Testing Of Preimplantation Embryosmentioning

confidence: 99%

The Historic Development and Incorporation of Four Assisted Reproductive Technologies Shaping Today′s IVF Industry

Schiewe¹

2016

JFIV Reprod Med Genet

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The concept of assisted reproductive technologies was most notably derived from the late 19th Century experiments of Sir Walter Heape who successfully transplanted rabbit embryos. Interestingly, it was not until the late 1940's and 1950's that renewed interests in rabbit embryo transfer and cryobiology occurred. The history behind developing effective procedures can be fascinating, though few could be more accidental than Dr. Chris Polge's discovery of glycerol (1948), from a mislabeled bottle of sugar solutions, being an effective cryoprotective agent for sperm freezing. The purpose of this review paper is to discuss four key scientific breakthrough technologies occurring between 1985 and 1995, which ultimately shaped the future of today's human in vitro fertilization (IVF) industry. More importantly, this paper highlights the foundation of underlying related discoveries and some unique stories involving their development and publication. In the end, this paper emphasizes the value of understanding scientific discovery timetables and the eventual re-discovery in the hands and minds of creative, determined and dedicated scientists, as history tends to repeat itself before its useful application is realized.

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“…The PCR can amplify single molecules of a target nucleic acid sequence sufficiently to permit isotopic [13][14][15][16] or, if the test sample contains little background DNA, non-isotopic detection [17][18][19][20][21][22] . PCR requires specialized equipment that is customized to fluctuate between specifically timed temperature variations.…”

mentioning

confidence: 99%

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2014

IJPSR

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This paper is specifically designed to explain the important scientific points of Polymerase Chain Reaction (PCR) technology in forensic science and molecular biology. Scientific procedures presented in the paper are complex, simple were specifically designed to better explain and reinforce the key concepts of PCR. The PCR is a laboratory technique for amplifying a specific sequence of a minute amount of Deoxyribonucleic acid (DNA) even from a single hair root or a microscopic blood stain left at a crime scene. Todays, PCR has found widespread and innumerable uses in forensics, genetics, DNA fingerprinting, bacteriology, virology, cloning and many other areas. Major research areas such as biomarker discovery, gene regulation and cancer research are challenging today's PCR technologies with more demanding requirements. Reproducibility of data and time to results are still major problems encountered by researchers.

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Diagnosis of Beta-Thalassaemia by Dna Amplification in Single Blastomeres From Mouse Preimplantation Embryos

Cited by 87 publications

References 7 publications

Direct electrophoretic detection of the allelic state of single DNA molecules in human sperm by using the polymerase chain reaction

Direct electrophoretic detection of the allelic state of single DNA molecules in human sperm by using the polymerase chain reaction

The Historic Development and Incorporation of Four Assisted Reproductive Technologies Shaping Today′s IVF Industry

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