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Introduction to Biotechnology Mr. Tsigaridis Why use it? What are some of the advantages to biotechnologies in genetics? The offer us advances in pharmacogenomics They allow individuals to identify problems early on They allow for identification of changes at the genetic level Why use it? What are some of the disadvantages to biotechnologies in genetics? They create ethical issues (euthanasia, abortions) WE DON’T KNOW What is out there? The biotechnologies that we are going to look at in this presentation are as follows: Amniocentesis Fetoscopy Ultra Sound Chorionic Villus Sampling PCR SDS-PAGE Adenovidus Vectoring (gene therapy) DNA Micro Array Amniocentesis What is amniocentesis It is a medical procedure used in prenatal diagnostics A small amount of amniotic fluid is sampled The fluid contains fetal tissue It is extracted through the amnion (sac) DNA is then examined for abnormalities Amniocentesis Amniocentesis Application Amniocentesis Application What can it be used for? It can be used to identify the sex of a fetus It can be used to identify genetic disorders, either from karyotyping or from genetic analysis (mapping of genome) Is this good or bad (talk about it with a partner, and then bring your ideas back to me) Some of the dangers Infection can occur from the puncture Incomplete healing can cause leakage and infection of the amniotic sac Fetoscopy What is a fetoscopy This is an endoscopic procedure that allows the medical professional access to the fetus, amniotic cavity, the umbilical cord, and the fetal side of the placenta Fetoscopy What is it used for? It allows for medical interventions such as: Biopsy Laser occlusion of damaged blood vessels Fetoscopy An important use It can also be used to perform blood transfusions of fetuses It can save the fetuses life It can also be a first step in identifying genetic disorders that a fetus may have Dangers of fetoscopies The incisions may become infected Leakage of the amniotic fluid into the body cavity may occur Chorionic Villus Sampling Is a form of prenatal diagnostic testing It is used to identify prenatal chromosomal and genetic disorders How it is done? A sample of the chorionic villus is taken and tested What is the chorionic villus? Placental Tissue When it is done? Its advantage comes with its timing It can happen from weeks 10-13 after the last period This is a better technique than amniocentesis because that can only happen from weeks 15-18 CSV – Chorionic Villus Sampling Reasons for having it done Mother's age of 35 years or greater Abnormal first trimester screen results Increased nuchal translucency or other abnormal ultrasound findings Family history of a chromosomal abnormalities or other genetic disorders Parents are known carriers for a genetic disorder UltraSound Is cyclic sound pressure with a frequency that is greater than that of human hearing UltraSound The production of ultrasound is used in many different fields It is used to penetrate a medium and measure the reflection signature This reveals information about the inner structure of the medium The old ultrasound The new ultrasound Other uses Studying the earth Cleaning teeth Breaking kidney/gull stones PCR – Polymerase Chain Reaction It is widely used in molecular biology Created by Kary Mullis This allows for amplification of small amounts of DNA The secret Thermus Aquatiucs AKA - TAQ Why Taq? The reason they wanted to use this bacteria was because it lives at such high temperatures The Polymerase (the enzyme that lays down nucleotides) works at high temperature Higher than that of DNAs denaturing point The process What goes in the tube? DNA of interest Primers Taq Polymerase Nucleotides (lots of them) Water Magnesium Then what happens They put the eppendorf tubes into the PCR cycler The process is repeated many times Why it works? Some Math 20 = 1 21 = 2 22 = 4 23 = 8 223 = 8,388,608 236 = 68,719,476,736 What are the uses Studying genes Forensics Today – basically all gene related research SDS-PAGE Sodium dodecyl sulfate polyacrylamide gel electrophoresis, is a technique used in biochemistry, genetics and molecular biology to separate proteins according to their electrophoretic mobility They are separated as a function of their size What is it used for? Genetic testing Paternity testing Forensics The Apparatus The Point DNA Micro Array A DNA microarray is an arrayed series of microscopic spots of DNA Oligonucleotides Each spot contains picomoles of a specific unique sequence, such a stretch of a gene sequence What is a picomole? The breakdown pico p 10-12 nano n 10-9 micro µ 10-6 milli m 10-3 centi c 10-2 deci d 10-1 kilo k 103 mega M 106 giga G 109 tera T 1012 What is it used for It is used to do many thousands of experiments at once It can be used to identify if a certain gene is present Or it can be used to identify if a gene is expressed, over expressed or underexpressed What it looks like Gene Therapy – Adenovirus Vector Viral vectors are a tool commonly used by molecular biologists to deliver genetic material into cells A common virus is adenovirus, HSV