Lab Reviews p GLO Lab Purpose of the

Lab Reviews

p. GLO Lab • Purpose of the Lab: – Genetic Transformation: take a gene from jellyfish Aequorea victoria and insert it into a bacteria. • Hypothesis: – If the bacteria expresses the jellyfish gene that produces the fluorescent protein, then it will glow green under UV light

p. GLO Lab • Understanding the concept: – Move gene from one organism to the other using a plasmid – Plasmid codes for GFP and a gene resistant to antibiotic ampicilin – Plasmid: gene regulation system, control expression of gene – Arabinose: sugar that when paired with plasmid containing GFP = glowing!

p. GLO Lab • Results: – Plate 1: +p. GLO, LB/amp • Color was white • Colonies present – Plate 2: +p. GLO, Colonies LB/amp/ara • Appear green under UV light • Colonies present – Plate 3: -p. GLO, LB/amp • No growth (why? ) – Plate 4: -p. GLO, LB • Colonies present • White in color

p. GLO • Discussion: – What is the genetic advantage of being able to turn on and off genes? – Why was ampicilin resistance critical for the growth of bacteria? – What was the role of arabinose?

Crime Scene Investigator • Purpose of the Lab: – Using PCR and agarose gel electrophoresis to analyze DNA samples of four suspects. Identify the person that committed the crime. • Hypothesis: – If the genetic marker of a suspect matches the crime scene DNA, then the individual who conducted the crime will be revealed.

Crime Scene Investigator • PCR: – Polymerase Chain Reaction – Produces large amounts of a specific piece of DNA from little amounts of starting (template) DNA – Need one template strand present, reaction buffers, four DNA subunits, DNA polymerase, DNA primers (MASTERY Mix) • Know why you use each of the following – Buffers: aid the enzyme – Primers: complementary to DNA strand needed because DNA polymerase cannot create a new DNA chain on its own (it needs preexisting chains of nucleotides)

Results

Results • STRS – Short Tandem Repeats • Repeated allele forms that will be present on the gel • Gel Electrophoresis – Bands move according to size (length) – Negative to positive

GMO Investigation • Purpose: – Extract DNA from food samples, run PCR analysis to amplify GMO potential sequence from the DNA and electrophorese the samples • Hypothesis: – If there is a 200 bp band from test food and GMO primers then the sample is GMO.

Procedure Clarifications: GMO • Was DNA extraction successful? – 2 sets of primers • Red: detect GMO-specific sequences – 200 – 250 bp • Green: plant DNA, detect GMO-derived or not – Allows you to tell if a GMO-negative result is due to lack of GMO material or unsuccessful DNA extraction – Amplify a 455 bp

GMO Lab Results • Lane 4 – If a band is present around 200 bp then GMO is present! • Has the typical GMO sequence present in foods that have been modified