GiM Flashcards
Southern Blotting
Used for large amounts of DNA
Northern Blotting
Used for RNA
Western Blotting
Used for proteins
PCR
Determines presence/absence (allele-specific) or product size
QF-PCR
Amplifies repeat using fluorescent primers
Products separated by size of repeats
Used in amniocentesis+ chorionic villi biopsy
qPCR
Confirms small copy number variations between pt and control
Oligonucleotide Ligation Assay
Compares 2 alleles with slight differences
Distinguishes between a disease-causing mutation and a normal allele
Sequencing
Sanger= type and place of mutation Clonal= cheaper and faster, used for PCR product, mutated genes and exome panels
FISH
Chromosomes labelled with dye
Identifies copy number imbalances, aneuploidy, cancer
Multiplex Ligation-Dependent Probe Amplification
DNA based PCR
Identifies copy number variations in many loci (right number of DNA)
Microarray CGH
Genome+control DNA compete Does 8 pts at a time Finds genetic imbalances Database identifies pathology of imbalances Needs good quality DNA
Next Generation Sequencing
Similar to microarray CGH with molecular barcode
>1= gain
Familial Inheritance
λ𝑠=𝑟𝑖𝑠𝑘 𝑜𝑓 2𝑛𝑑 𝑠𝑖𝑏𝑙𝑖𝑛𝑔/𝑟𝑖𝑠𝑘 𝑜𝑓 𝑔𝑒𝑛𝑒𝑟𝑎𝑙 𝑝𝑜𝑝𝑢𝑙𝑎𝑡𝑖𝑜𝑛
Knudson’s Two Test Hypothesis
Need two mutations in cancer genes to make a tumour
Need 3 generations to say cancer is familiar
e.g. retinoblastoma, FAP, HNPCC, BRCA, Li-fraumeni (p53)
Non-Mendelian
Additive genes e.g. BP, head circumference, weight
Influenced by environ
Diseases= spina bifida, asthma, cancer, DM
Androgenesis
No egg in ovum but fertilised by sperm
Makes hydatiform mole (malignant trophoblastic tumour)
Well developed membranes
Parthenogenesis
Two eggs in ovum+no sperm Benign ovarian teratoma Mostly epithelium (no muscle, membranes or placenta)
Imprinting
Epigenetic
Maternal or paternal copy methylated (silencing it at CG island)
Hypomethylation= Russell-Silver syndrome
Angelman
Deletion in chr 15 on MATERNAL gene
Dysmorphic, seizures, jerky, mental handicap
Prader-Willi
Deletion in chr 15 on PATERNAL gene
Obese, mental handicap, hypergenitalism, hypotonia
X-Inactivation
One X silenced in females
In males, X or Y silenced
Occurs in blastocyst
Cytogenetics
Found via G-banding, FISH or qPCR
Issues= dosage, genes activated/inactivated, gene position changed, unmasking of recessive
Aneuploidy
Increased maternal age increases risk as meiotic structure deteriorates
Edward’s
Trisomy 18
Microcephaly, clenched hands, mental delay, congenital heart disease
Patau
Trisomy 13
Like edwards but also brain defects, cleft lip, polydactyly, abnormal genitalia
Turner’s
X
No puberty
Short, reduced IQ, aortic coarctation
Klinefelter’s
XXY
Testicular dysgenesis, tall, long limbs, low IQ
Polyploidy
Gain of whole sets of chromosomes
Triploidy caused by digyny, diplospermy or dispermy
Mosaicism
Caused by mitotic non-dysjunction
Errors at cleavage
Reciprocal Translocations
Break+ exchange
Only phenotype risk if a gene is broken
Balanced
Robertsonian Translocation
Whole arm fusion
Acrocentric chromsomes (13,14,15, 21,22)
No phenotype risk
Balanced
Inversion
2 breaks with rotation and rejoining
Pericentric or paracentric
Balanced
Insertion
Onto another chromosome
Balanced
Interstitial Deletion
In middle of chromosome
Terminal Deletion
Off the end of a chromosome
Can form a ring
Unbalanced Rearrangments
Copy number variation
Detected for FISH, MPLA, Microarray CGH, NGS, qPCR
Genes
Have exons, introns and regulatory sequences
Repetitive DNA can be in satellites or interspersed
Exon Skipping
Transcribing can skip an exon between two others
Mutually Exclusive Exon Choice
Skipping a small exon if it is next to another small exon
Ancestral Genes
Duplicated and dispersed amongst genome
Some non functional= pseudogenes
William’s
7q11 deletion Hypercalcaemia, cocktail party speech, learning problems Heart problems (supravalvular aortic stenosis and peripheral pulmonary artery stenosis)
Huntingdon’s
36+ CAG repeats at HTT gene
More repeats in later generations
Beckwith-Wiedermann
Caused by hypermethylation (imprinting)
Risk of Wilms tumour
Achondroplasia
AD
Limb shortening
2q11-2 Deletion
Variable symps
Most have heart defects
Pharmacogenetics
Drug response altered by SNPs, gene amplification, translocations, deletions and insertions