JeopardyPCR Gel

ElectrophoresisMisc. PV92 and

morepGLO and

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Misc

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PCR is possible due to the discovery and use of thermostable polymerases. Why was this

crucial to the PCR process?

The steps of PCR run at much higher temperatures than occur in most living

organisms

Why do forensic labs analyze non-coding DNA and not coding areas of a chromosome?

Non coding areas can accumulate mutations that do not negatively

affect organisms?

What are dNTPs?

Free nucleotides, sources of A, T, G, and

C in a PCR reaction

Hybridization of a primer with its complement on the template…. Give term

Annealing

Replication of DNA many times… give term

Extension

Toward which pole (positive or negative) does DNA migrate when electric current is run through the gel? Why?

Positive, because DNA is negatively charged

Give three uses for DNA analysis

Forensic fingerprintingPaternity testing

Evolutionary relationships

Give one drawback of RLFP analysis

Requires a large sample

Lt. Russ is investigating a murder scene. The felon was scratched by his victim & some of his skin cells were found under the victim’s fingernails.

A DNA test was performed. Which of the suspects is the murderer?

Suspect 2

Mr. & Mrs. Jones just gave birth to fraternal twins- Bob and Jane. Unfortunately, the nurse has confused the Jones twins with 4 other babies.

The doctors took samples of DNA from each of the babies and Mr. & Mrs. Jones. Which of the 6 children are Mr. & Mrs. Jones twins?

What is 1 & 5?

What would happen during electrophoresis if the gel concentration was too low for a specific sample.

Samples would run off or not separate

because the holes in the matrix would be

too large.

How many amplified loci are used to establish a unique DNA fingerprint for suspects in a

crime ?

13

What would happen if you ran the gel electrophoresis without first using SDS on the

protein when separating proteins?

The proteins would have different charges and not

separate by size correctly?

The number of amplified pieces of DNA equals _______ after 7 cycles of PCR

128

What is isoelectric focusing?

Separation of proteins by creating stable pH

gradients in a gel and using the variabilty of

protein charges as separation criteria?

What cellular structures must be broken down to release the DNA from a cell?

The cell and nuclear membranes must be broken to release the

template DNA

into the solution.

What kind of controls are run in the PV92 experiment? Why are they important? Could

others be used?

The controls that are run in this experiment are the homozygous +/+, homozygous –/–, and heterozygous +/– known samples. These bands

have known base-pair lengths and can be used in comparison to unknown student samples.

This shows a class run of PV92, What is in lane 2, 3, and 4

Standards, two homozygous and one heterozygous

This shows a class run of PV92, how many students are heterozygous?

6 - #’s 6,10,13,18,19,&20

What is the difference between an allele and a locus?

Locus is location on chromosome

Allele is the version of gene present on that locus

What kinds of changes to chromosomes would cause a "polymorphism" to appear in the DNA profiles of

different individuals from a population?

A mutation or rearrangement could cause a primer site to be lost, causing a band to disappear.

An insertion or a deletion between primer sites would cause the band to migrate a shorter or longer distance,

respectively.

In the pGLO lab the source of the In the pGLO lab the source of the fluorescent gene of interest is?fluorescent gene of interest is?

Jellyfish

What compound is required in an agar plate to make the transformed bacteria glow?

Arabinose

Why does the plasmid have to be inserted into a cell to make the glow?

It is the protein that glows and only cells with the “glow” instructions can pass them on to ribosomes that make proteins!

Inserting a new gene into an organism is called________.

Transformation

Why don’t bacteria destroy their own DNA with their restriction enzymes?

Methylation!!

In the pGLO lab what 3 methods or materials are used to increase transformation efficiency?

Heat ShockCold Shock

Salts

To visualize DNA fragments after electrophoresis you must then….

Stain the gel with an appropriate stain like

Fast Blue

Why do you run molecular weight Why do you run molecular weight standards in one lane during standards in one lane during electrophoresis?electrophoresis?

To determine the relative sizes of the DNA sample

How many more days of school are left ?

…… It depends on who is

counting and when you play this Jeopardy game

……..