Week 10 Lab Review

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DNA's Charge

DNA is negatively charged due to the phosphate backbone. This means it will be repelled by negative charge, and attracted by positive charge. NOTE: Since DNA molecules all have the same charge, the migration rate is solely based on their size!

DNA Ladders: Goal

Determine the size of unknown DNA fragments by running gel electrophoresis on them with a corresponding ladder.

Electrophoresis

Is the study of the movement of charged particles in an electric field.

Gel Electrophoresis

It is a technique that separates macromolecules according to their size and charge. The overall charge of a molecule affects the speed at which it travels through the gel Highly charged molecules migrate more quickly through the gel than weakly charged molecules. The mobility of a molecule also depends on its molecular size, shape. The small pores of the gel matrix act as a sieve that provides great resolving power

DNA Ladders: Ladder

Known fragments of DNA are mixed together for use as a standard curve for each gel electrophoresis run.

Characteristics involved in Gel Electrophoresis

Small molecules are able to more easily through the pores than larger molecules and so travel more quickly Large molecules encounter more resistance as they travel through the tiny pores and so travel more slowly Size and net charge are factors that together determine how quickly molecules will travel through the gel and ultimately what their migration distance will be. Small size and strong charge INCREASE a molecules migration rate through the gel Large size and weak charge DECREASE the migration rate

Agarose

a purified form of agar, a gelatinous substance extracted from red algae

Parts of Gel Electrophoresis: Running buffer

keeps gel cool, and conducts electric field through gel

Parts of Gel Electrophoresis: Power Supply

provides constant voltage electric field

Parts of Gel Electrophoresis: Gel rig with electrodes

provides electric field to move DNA

Parts of Gel Electrophoresis: Gel

serves as matrix for DNA to move through


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