Introduction to Biotechnology

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Introduction to Biotechnology

• Mr. Tsigaridis

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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

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Why use it?

• What are some of the disadvantages to
  biotechnologies in genetics?
• They create ethical issues (euthanasia,
  abortions)
• WE DONT KNOW

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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

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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

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Amniocentesis
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Amniocentesis Application
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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)

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Some of the dangers

• Infection can occur from the puncture
• Incomplete healing can cause leakage and
  infection of the amniotic sac

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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

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Fetoscopy

• What is it used for?
• It allows for medical interventions such as
• Biopsy
• Laser occlusion of damaged blood vessels

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Fetoscopy
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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

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Dangers of fetoscopies

• The incisions may become infected
• Leakage of the amniotic fluid into the body
  cavity may occur

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Chorionic Villus Sampling

• Is a form of prenatal diagnostic testing
• It is used to identify prenatal chromosomal and
  genetic disorders

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How it is done?

• A sample of the chorionic villus is taken and
  tested
• What is the chorionic villus?
• Placental Tissue

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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

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CSV Chorionic Villus Sampling
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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

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UltraSound

• Is cyclic sound pressure with a frequency that is
  greater than that of human hearing

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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

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The old ultrasound
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The new ultrasound
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Other uses

• Studying the earth
• Cleaning teeth
• Breaking kidney/gull stones

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PCR Polymerase Chain Reaction

• It is widely used in molecular biology
• Created by Kary Mullis
• This allows for amplification of small amounts of
  DNA

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The secret

• Thermus Aquatiucs
• AKA - TAQ

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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

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The process

• What goes in the tube?
• DNA of interest
• Primers
• Taq Polymerase
• Nucleotides (lots of them)
• Water
• Magnesium

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Then what happens

• They put the eppendorf tubes into the PCR cycler
• The process is repeated many times

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Why it works?
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Some Math

• 20 1
• 21 2
• 22 4
• 23 8
• 223 8,388,608
• 236 68,719,476,736

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What are the uses

• Studying genes
• Forensics
• Today basically all gene related research

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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

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What is it used for?

• Genetic testing
• Paternity testing
• Forensics

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The Apparatus
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The Point
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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?

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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

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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

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What it looks like
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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

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