We think you have liked this presentation. If you wish to download it, please recommend it to your friends in any social system. Share buttons are a little bit lower. Thank you!
Presentation is loading. Please wait.
Published byKayla Scott
Modified over 3 years ago
Chapter 29 Essential Concepts in Molecular Pathology Companion site for Molecular Pathology Author: William B. Coleman and Gregory J. Tsongalis
Companion site for Molecular Pathology Copyright © 2009 by Academic Press. All rights reserved. 2 FIGURE 29.1 Schematic representation of the Ce\GG repeat in exon 1 of FMR1 and associated alleles. A CGG-repeat number less than or equal to 45 is normal. A CGG-repeat number of 46 to 54 is in the gray zone and has been reported to expand to a full mutation in some families. A CGG-repeat number of 55 to 200 is considered a premutation allele and is prone to expansion to a full mutation during female meiosis. A CGG-repeat number in excess of 200 is considered a full mutation and is diagnostic of fragile X syndrome.
Companion site for Molecular Pathology Copyright © 2009 by Academic Press. All rights reserved. 3 FIGURE 29.2 Southern blot analysis for the diagnosis of fragile X syndrome. Patient DNA is simultaneously digested with restriction endonucleases EcoR1 and Eag1, blotted to a nylon membrane, and hybridized with a 32P-labeled probe adjacent to exon 1 of FMR1 (see Figure 29.1). Eag1 is a methylation-sensitive restriction endonuclease that will not cleave methylated DNA. Normal male control DNA with a CGG-repeat number of 22 on his single X chromosome (lane 1) generates a band about 2.8 kb in length corresponding to Eag1-EcoR1 fragments (see Figure 29.1). Normal female control DNA with a CGG-repeat number of 20 on one X chromosome and a CGG-repeat number of 25 on her second X chromosome (lane 5) generates two bands, one at about 2.8 kb and a second at 5.2 kb. EcoR1-EcoR1 fragments approximately 5.2 kb in length represent methylated DNA sequences characteristic of the lyonized chromosome in each cell that is not digested with restriction endonuclease Eag1. DNA in lane 2 contains an FMR1 CGG-repeat number of 90 and is characteristic of a normal transmitting male. The pattern observed in lane 3 is representative of a mosaic male with a single X chromosome with a full mutation (>200 repeats). However, the full mutation in some cells is unmethylated; in other cells, the full mutation is fully methylated, hence the term mosaic. In those cells in which the full mutation is unmethylated, digestion by both Eag1 and EcoR1 occurs, and in those cells in which the full mutation is fully methylated, digestion of the DNA by Eag1 is inhibited. The pattern observed in lane 4 is diagnostic of a male with fragile X syndrome, illustrating the typical expanded allele fully methylated in all cells. Lane 6 is characteristic of a female with one normal allele that has 29 CGG-repeats and a larger gray zone allele with a CGG-repeat number of 54. Lane 7 is the pattern observed from a premutation carrier female with one normal allele having a CGG-repeat number of 23 (band at about 2.8 kb) and a second premutation allele with CGG repeats of 120 to about 200 (band at about 3.1 kb). In premutation carrier females, in cells in which the X chromosome with the premutation allele is lyonized, the normal 5.2 kb EcoR1-EcoR1 band is larger because of the increased CGG-repeat number and is about 5.5 kb in length. Lane 8 is diagnostic of a female with fragile X syndrome with one fully expanded mutation allele that is completely methylated and a second normal allele with a CGG-repeat number of 33.
Companion site for Molecular Pathology Copyright © 2009 by Academic Press. All rights reserved. 4 FIGURE 29.3 HCV genotyping assay. Comparison of samples from patients infected with HCV genotypes 1b, 2a/c, 2b, and 3a. Shown are genotype-specific melting transitions for four samples in 2 mM MgCl2. Data were obtained by monitoring the fluorescence of the LCRed640-labeled FRET sensor probe during heating from 40 to 80°C at a temperature transition rate of 0.1°C/s.
Companion site for Molecular Pathology Copyright © 2009 by Academic Press. All rights reserved. 5 FIGURE 29.4 T-cell receptor gamma chain PCR assay for clonality. (A) Polyclonal reactive T-cell proliferation pattern. A polyclonal population of T-cells with randomly rearranged T-cell receptor gamma chain genes produces a normal or Gaussian distribution of fluorescently labeled PCR products from each primer pair in the multiplex reaction. This produces four bell-shaped curves that represent the valid size range for an individual primer pair. Two of the valid size ranges are green and two are blue; G1, B1, G2, B2. The red peaks represent size standards. (B) Clonal T-cell proliferation pattern. A clonal T-cell proliferation results in a relative dominance of a single T-cell receptor gamma chain gene producing a predominant spike of a discrete size on the corresponding electropherogram. Data were obtained using an ABI 3100 capillary electrophoresis system and ABI Prism Software.
Chapter 6 Essential Concepts in Molecular Pathology Companion site for Molecular Pathology Author: William B. Coleman and Gregory J. Tsongalis.
Chapter 12 Essential Concepts in Molecular Pathology Companion site for Molecular Pathology Author: William B. Coleman and Gregory J. Tsongalis.
Introduction Sandeepa Chauhan, Madhumita Roy Chowdhury, Neerja Gupta, Sheffali Gulati*, BK Thelma #, Anjali Dabral #, Madhulika Kabra Genetic Unit, Department.
Chapter 6 PART II: Concepts in Molecular Biology and Genetics The Human Genome: Implications for the Understanding of Human Disease Companion site for.
Problem 1 Among 4.5 million births in one population during a period of 40 years, 41 children diagnosed with the autosomal dominant condition aniridia.
Diagnosis with PCR This is a preparation of DNA. We zoomed in a portion of a gene. We know that two primers, Forward and Reverse, will hybridize at specific.
Fragile X Syndrome.
Chapter 31 Essential Concepts in Molecular Pathology Companion site for Molecular Pathology Author: William B. Coleman and Gregory J. Tsongalis.
DNA Biochemical Processes and Forensic Applications. 1.
SP5 -Validation of Methylation Specific PCR for the detection for large FMR-1 expansion mutations in males and females Stacey Mutch East Midlands Regional.
Plant Breeding Shree Krishna Adhikari ©Shree Krishna Adhikari.
Discuss the relationship between phenotype and genotype in Fragile X syndrome. Louise Williams 06/03/2008.
Biotech. Southern Blotting Through a series of steps, DNA that has been separated by electrophoresis is applied to a membrane of nylon or nitrocellulose.
Manipulating DNA Genetic Engineering uses the understanding of the properties of DNA to study and change DNA sequences in living organisms – Invitro… in.
Investigating the use of Multiple Displacement Amplification (MDA) to amplify nanogram quantities of DNA to use for downstream mutation screening by sequencing.
ABC for the AEA Basic biological concepts for genetic epidemiology Martin Kennedy Department of Pathology Christchurch School of Medicine.
DNA Mutations and Repair Chapter 18 Part 1. Gene Mutations and Repair Nature of mutations Causes of mutations Study of mutations DNA repair.
Chapter 20 DNA Technology and Genomics. Biotechnology is the manipulation of organisms or their components to make useful products. Recombinant DNA is.
Restriction Fragments and Mapping Restriction Fragment Analysis – System used to compare the genes and DNA sequences between individuals in a population.
Chapter 5: Exploring Genes and Genomes Copyright © 2007 by W. H. Freeman and Company Berg Tymoczko Stryer Biochemistry Sixth Edition.
Biotech. Cloning a mammal PCR This is the polymerase chain reaction. It is a technique to multiply a sample of DNA many times in a short period of time.
Chapter 17: Variable-Number Tandem Repeats Profiling.
Locating and sequencing genes
Types of STR markers- 5 types based on sequence STR allele nomenclature Allelic ladder Serological methods of identity profiling Identity profiling.
Forensic Biology by Richard Li
Molecular Biology of Genes Chapters DNA Technology (not in your book)
Simple-Sequence Length Polymorphisms SSLPs Short tandemly repeated DNA sequences that are present in variable copy numbers at a given locus. Scattered.
Synteny - many distantly related species have co- linear maps for portions of their genomes; co-linearity between maize and sorghum, between maize and.
DNA: Review, Replication, & Analysis Two types of DNA Nucleic DNA –Found in the nucleus of a cell –Specific to an individual Mitochondrial DNA (mtDNA)
DNA fingerprinting Every human carries a unique set of genes (except twins!) The order of the base pairs in the sequence of every human varies In a single.
DNA basics DNA is a molecule located in the nucleus of a cell Every cell in an organism contains the same DNA Characteristics of DNA varies between individuals.
Chapter 5 Nucleic Acid Hybridization Assays A. Preparation of nucleic acid probes: 1. Labeling DNA & RNA - Nick Translation - Random primed DNA labeling.
DNA (gene mutations, paternity, organs compatibility for transplantations) RNA Proteins (gene expression)
Molecular Basis for Relationship between Genotype and Phenotype DNA RNA protein genotype function organism phenotype DNA sequence amino acid sequence transcription.
© 2013 Pearson Education, Inc. Extensions of Mendelian Genetics Incomplete Dominance is when a heterozygote expresses a phenotype intermediate between.
6.3 Advanced Molecular Biological Techniques 1. Polymerase chain reaction (PCR) 2. Restriction fragment length polymorphism (RFLP) 3. DNA sequencing.
Probes and screening for disease. Background Genetic disorders are often the result of gene mutations. People with a mutant allele often have a family.
Faculty of Science and Engineering Human Molecular Genetics
Kinship DNA Fingerprinting Simulation Grab the packet from the front table and begin reading.
Amplification of Genomic DNA Fragments OrR. Amplification To get particular DNA in large amount Fragment size shouldn’t be too long The nucleotide sequence.
Chapter # - Chapter Title $100 $200 $300 $400 $500 $100$100$100 $200 $300 $400 $500 Human Heredity Human Chromosomes Human Molecular Genetics Human Heredity.
Current Genetic Techniques How can we use DNA today? Section 3 - Parts of Chapters 13 & 14.
MOLECULAR DIAGNOSTICS IN CONGENITAL ADRENAL HYPERPLASIA (21- HYDROXYLASE DEFICIENCY)
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display Human Genetics Concepts and Applications Seventh Edition.
Problem 1 Mutation in noncoding sequence change the number of protein molecules produce, but, generally each protein molecule made will have a normal amino.
What is restriction fragment analysis? Restriction fragment analysis is a process used to compare the DNA of two or more different organisms.
Putting it all together: Finding the cystic fibrosis gene Cystic fibrosis (CF) is a genetic disorder that is relatively common in some ethnic groups A.
Chapter 10: Genetic Engineering- A Revolution in Molecular Biology.
Molecular Analysis of Genes and Gene Products BIT 220 Chapter 22.
Chapter 19 – Molecular Genetic Analysis and Biotechnology
© 2017 SlidePlayer.com Inc. All rights reserved.