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Lecture 11

HMB321H1 Lecture Notes - Lecture 11: Krabbe Disease, Molecular Genetics, Chromatin


Department
Human Biology
Course Code
HMB321H1
Professor
Maria Papaconstantinou
Lecture
11

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2 MARCH 2011
HMB321 L11: Introduction to Molecular Genetics
*** Information from the book is on the final exam (i.e. Please finish that stupid book)
-Focus on:
-DNA sequencing methodologies (454, Sanger,
-Big names in DNA sequencing (Venter, Chan, Steffanson,
What is molecular genetics?
-Study of processes regarding the replication, translation of DNA into further products
Inactive DNA: Chromatin
-DNA is wrapped around nucleosome => wrapped into further chromatin structures
-Noncoding genes have more regulatory roles vs. Coding genes (usually proteins or products)
Selected major breakthroughs
-Inheritance (Mendel)
-Linkage (Morgan)
-Helix
-See slides
Molecular Genetics & Disease
-Genetic diseases, e.g. Krabbe disease
-Knowing the sequence of gene can allow us to learn the function (genotype => phenotype
association), i.e. GALC -/- recessive
-Effects = less myelin production = lower development of baby
-Affliction displays low muscle tone
-Molecular genetics of viruses + other disease causing agents
-Chromosomal aberrations, e.g. Trisomy 21 (via nondisjunction or chromosomal translocations)
Isolation of DNA
-Homogenize tissue => lyse cells => remove proteins/RNA/debris ===> isolation ===> purification
PCR of DNA
-Primers anneal at a specific temperature (Tm) that are specific to the length/composition of primer
-Extension is often around 2min long (require primers, Taq polymerase + template strand of
DNA with DNTPs to create new strand) -- repeated several times with lag times in between
to ensure that the elongation process is being carried out to its necessary length
RT-PCR (Real Time: Quantitative)
-A way of counting how much products are produced
-Done via annealing a probe to your target sequence
-Primer displaces the probe; the probe’s promoter is displaced into the medium = allows
fluorescence
-The more PCR occurs = more broken probe reporters = quantify (via light analysis) how much
PCR made via how much fluorescence is found in media
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