Développement de méthodes d'analyse de l'ADN par clivage d'une chimère ARN/ADN et par spectrométrie de masse MALDI-TOF PDF Download
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Author: Florence Mauger
Publisher:
ISBN:
Category :
Languages : en
Pages : 345
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Book Description
As the DNA sequence of the human genome is now completed and available, interest is shifting towards the detection of DNA in candidate genes of human diseases. These DNA modifications can be either in the sequence or epigenetic. Thus, rare, frequent, known and unknown polymorphisms can be identified through development of new methods of sequencing for each individual. We have developed a new method for DNA analysis using cleavage of RNA/DNA chimera and MALDI-TOF MS. The method takes advantage of a novel class of thermostable DNA polymerases that also incorporate ribonucleotides. The RNA/DNA chimera contains three deoxyribonucleotides and the fourth nucleotide in its ribonucleotide form. These products can be either single-stranded linear or double-stranded. The RNA/DNA chimera is cleaved by sodium hydroxide after each ribonucleotide base. All fragments contain only one ribonucleotide 3’ terminal base with a phosphate terminal group. Fragments are desalted by the addition of cation exchange resin and masses are determined by MALDI-TOF MS. The mass fingerprint is used to identify deviations from the reference sequence. Gas phase fragmentations of the RNA/DNA chimera were also studied. With the high throughput capability of MALDI-TOF MS, this facile method is well suited for screening DNA sequences of limited genomic regions of interest in a large number of individuals. This rapid and accurate, new method, has many potential applications including: DNA genotyping, DNA multiplex allele-specific genotyping, DNA micro-haplotyping, DNA methylation analysis and DNA sequencing
Author: Florence Mauger
Publisher:
ISBN:
Category :
Languages : en
Pages : 345
Get Book Here
Book Description
As the DNA sequence of the human genome is now completed and available, interest is shifting towards the detection of DNA in candidate genes of human diseases. These DNA modifications can be either in the sequence or epigenetic. Thus, rare, frequent, known and unknown polymorphisms can be identified through development of new methods of sequencing for each individual. We have developed a new method for DNA analysis using cleavage of RNA/DNA chimera and MALDI-TOF MS. The method takes advantage of a novel class of thermostable DNA polymerases that also incorporate ribonucleotides. The RNA/DNA chimera contains three deoxyribonucleotides and the fourth nucleotide in its ribonucleotide form. These products can be either single-stranded linear or double-stranded. The RNA/DNA chimera is cleaved by sodium hydroxide after each ribonucleotide base. All fragments contain only one ribonucleotide 3’ terminal base with a phosphate terminal group. Fragments are desalted by the addition of cation exchange resin and masses are determined by MALDI-TOF MS. The mass fingerprint is used to identify deviations from the reference sequence. Gas phase fragmentations of the RNA/DNA chimera were also studied. With the high throughput capability of MALDI-TOF MS, this facile method is well suited for screening DNA sequences of limited genomic regions of interest in a large number of individuals. This rapid and accurate, new method, has many potential applications including: DNA genotyping, DNA multiplex allele-specific genotyping, DNA micro-haplotyping, DNA methylation analysis and DNA sequencing
Author: Mark S. Meskin
Publisher: CRC Press
ISBN: 1420031694
Category : Medical
Languages : en
Pages : 221
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Book Description
Are soy isoflavones neuroprotective? Just how different is one species of Echinacea from another? Which phytochemicals will be effective as therapeutic agents in vivo? Supported by solid scientific research, Phytochemicals in Nutrition and Health helps provide answers to these and other probing questions concerning the mechanisms of action associat
Author: Joginder Singh
Publisher: Springer Nature
ISBN: 9811528179
Category : Science
Languages : en
Pages : 377
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Book Description
Microbial biotechnology is an important area that promotes advanced research into using microbes for value-added products, human nutrition, and the overall wellbeing of society. This book presents the latest information on the use of microbes for sustainable development, and highlights state-of-the-art biotechnological techniques used to harness microbial biotechnological traits on a commercial scale. Gathering contributions from authoritative researchers in the field, it addresses recent advances in microbial biotechnological approaches that offer sustainable options for future generations. Exploring a broad range of microbial products and their uses, the book specifically places emphasis on the application of microorganisms in healthcare, the environment and industry. It also discusses various compound classes derived from microbial metabolites. Pursuing a holistic approach to recent advances in the utilization of various microbes as biotechnological tools, the book also covers traditional uses, and explores emerging strategies to harness their full potential. Accordingly, it offers a valuable resource for researchers and graduate students alike.
