Lab 7 Flashcards
Traditional Biotechnology example
selective plant and animal breeding
Modern Biotechnology
manipulation of DNA in vitro
Forensic Biology
scientific method use in legal and investigation
Samples must be _________ to provide DNA for analysis
Nucleated
Samples suitable for DNA analysis
blood, hair, cheek, bodily fluids,
Steps to analyse DNA
1: DNA extraction
2: Polymerase Chain Reaction (PCR)
3: Restriction Fragment Analysis
4: Interpretation
DNA extraction explaination and ingredients
chemically lysing cells to liberate DNA.
-Isotonic Medium
-Lysis Solution
-Ethanol
Step one of DNA extration
Putting the sample into an isotonic solution so they do not shrink or burst
Step two of DNA extraction
Adding a lysis solution to break bonds in lipid/protein membranes to get out DNA without killing it
Step three of DNA extraction
Adding ethanol to make DNA precipitate because it is highly soluble in water but not in ethanol
Four ingredients of PCR
DNA extract
Deoxyribonucleotide triphosphates (dNTP: dATP, dCTP, dGTP, dTTP)
Primers
Taq polymerase (heat stable enzyme to elongate DNA by adding dNTPs)
Steps to PCR
Denaturation of DNA
Annealing of Primers
Extension of Primers
Repetition until desired amount of DNA available
Denaturing of DNA
heating of DNA to break apart into single strands
Annealing of primers
sample cooled, primers attach to start dna replication
Extension of primers
sample heated slightly to allow Taq polymerase to attach and add nucleotides
Restriction Fragment analysis and steps
Cuts DNA regions out
-Restriction Digest
-Gel Electrophoresis
Restriction Digest
restriction enzyme added to the product and solution placed in an incubator. DNA cut into specific fragments
Gel Electrophoresis
Allows evaluating upon molecule size. Since DNA is negative it moves towards the positive electrode. smaller chunks will move farther than larger chunks
Purpose of ladder in Gel Electrophoresis
having a known sequence of DNA and being able to compare with known size of DNA
Does the prevalence of each allele vary by ethnic group
Yes
Is the pattern of restriction fragments in gel called a DNA footprint?
No
