Detection of Hereditary Breast Cancer Breast Cancer Genes Two genes associated with inheritance of breast cancer Breast cancer gene 1 (BRCA1) Chromosome 17 BRCA 2 Chromosome 13 When normal Both are tumor suppressor genes Repair damage and prevent cancer cells from forming When mutated
Lead to breast or ovarian cancer BRCA 1 and BRCA 2 5 to 10% of all breast cancers are because of these genes Women 12% chance of developing breast cancer in lifetime Goes up to 85% if said woman has mutated BRCA 1 or 2 If male has BRCA 2 mutation 1 in 10 ratio of getting breast cancer (100,000 jump) Family Members Judy is worried!!!!! Jennifer has DNA sequencing
Jennifer had breast cancer Test positive of BRCA 2 (negative for BRCA 1) Laura Tests positive for same mutation as Jennifer BRCA2 We will be looking at BRCA2 Contains 80,000 nucleotides 600 mutations associated with BRCA2 Most cause increased incidence of breast cancer (not all) Most of these mutations are insertion or deletion
Marker Analysis (Haplotyping) Due to expense of DNA sequencing We will use marker analysis to test Jennifer and Diana Marker analysis Genetic test Gene mutation is analyzed using a genetic marker Instead of analyzing genet itself Genetic marker: short sequence of DNA associated with a particular gene or trait with a known location on a chromosome
Short Tandem Repeats Genetic markers used in marker analysis are short DNA sequences Also called microsatellites STR = region of DNA composed of a short sequence of nucleotides repeated many times. Number of repeated STRs varies from person to person Different number of repeats = different alleles Most occur in introns (non-coding DNA) Do not affect gene function STR for BRCA 2
Location = chromosome 13 STR analysis for this lab It is on 13, next to BRCA 2 gene Gel Electrophoresis Different STRs have different repeats Gel will separate alleles based on number of repeats More repeats travels less Less repeats travels more Loading Samples
5 uL: 130 Volts for 30 minutes Lane #1: DNA Size Markers Lane #2: Helens DNA Lane #3: Harolds DNA Lane #4: Susans DNA Lane #5: Adams DNA Lane #6: Negative Control
Determining Size of DNA Fragment Using a ruler Measure (ON EACH BAND!!!!) Distance of DNA fragment from origin (gel well) Distance from the origin (gel well) to the tracking dye Calculate the Rf value Distance the DNA fragment has migrated from the origin (gel well) Rf = Distance from the origin (gel well) to the reference point (tracking dye) DNA Size Markers Fragment Length in Base Pairs
Fragment 1 1353 Fragment 2 1078 Fragment 3 872 Fragment 4
603 Fragment 5 310 Fragment 6 281 Fragment 7 234
Fragment 8 194 Distance Migrated (mm) A Distance to Reference Point (mm) B Rf AB
DNA Sample: Diana Fragment: Fragment 1 Fragment 2 Jennifer Fragment 1
Fragment 2 Laura Fragment 1 Fragment 2 Judy Fragment 1 Fragment 2
Distance Migrated (mm) A Distance to Reference Point (mm) B Rf AB DNA Sample: Fragment:
Diana Fragment 1 Fragment 2 Jennifer Fragment 1 Fragment 2 Laura
Fragment 1 Fragment 2 Judy Fragment 1 Fragment 2 Fragment Length (in base pairs) Allele Present:
Fragment Length in Base Pairs: Allele: 200 Allele 1 300 Allele 2 400
Allele 3 500 Allele 4 600 Allele 5 700 Allele 6
800 Allele 7 900 Allele 8 1000 Allele 9 Questions
1. Which allele is associated with the BRCA2 mutation? Explain your answer. 2. Which family members have the BRCA2 mutation? Explain your answer. 3. Explain whether you think Judys family occurrences of breast and ovarian cancers are sporadic, hereditary, or familial. 4. Is Judy a good candidate for BRCA1 or BRCA2 genetic testing? Explain your answer.
Ensure you know 'absolute' information: Irwin Mitchell Factsheets & Template letters IPSEA - on-line training Douglas Silas Brown Jacobson Always be absolutely open and transparent - engage parents/carers positively and pro-actively Keep the young person central to everything Remain resolute...
Introduction. Osteoarthritis is the . most prevalent . complex, degenerative chronic disease. It is a disorder of the hyaline joints characterized by wear, softening and thinning of the articular cartilage and diminished compliance of the sub-chondral bone.
Goal is totally distributed application. No point of failure. No censorship. Totally transparent . App Logic via smart contract. App data via decentralized storage like IPFS or Swarm. Frontend via IPFS or Swarm as well . Name resolution via ENS...
The two categories of software are system software and application software. System . Software. System software . consists of the programs that control or maintain the operations of the computer and its devices. System software serves as the interface between...
Keeyoon Sung, Linda R. Brown, Timothy J. Crawford. Jet Propulsion Laboratory, California Institute of Technology, ... of Physics and Astronomy, The University of Lethbridge, 4401 University Drive, Lethbridge, Alberta, Canada . co-authors are: Objective. Prior studies of the band investigated...
Arial MS Pゴシック Calibri Wingdings Trebuchet MS blank Parenting work in the youth justice system Supporting evidence base Youth Justice Board February 2010 Contents Slide 3 YOTs deliver a large number of interventions to parents and families in high need...
Unit 8: The lives of rich and poor people in Tudor times Differences between rich and poor people in Tudor times Rich Poor Houses Food Clothes Leisure Health Large, comfortable, nice furniture, glass windows Small, dark, often damp, very little...