Process: Gel Electrophoresis

Introduction:

Gel electrophoresis is done whenever you want to see the size and quantity of DNA. The materials involved in gel electrophoresis pose minimal risk, with one notable exception. Ethidium Bromide (EtBr) is used to stain the DNA, and is a carcinogen and a mutagenic material. Caution is necessary when handling this material. Gloves are essential, and the gel must be disposed properly in a designated waste container.

Equipment/Supplies needed:

Weigh boat

Analytical balance

250 ml Erlenmeyer Flask

8 tooth comb

Gel casting tray

Lab tape

Molecular weight ladder

Para film

Spatula

A 2 to 20 µl pipette

Loading dye

Power Supply

TAE buffer

Samples

Cautions:

Liquid gel may be hot. Handle with care DNA is a Bio-hazard and needs to be treated with extreme care according to Bio-

hazard safety regulations. The next steps involve the use of EthBr. EthBr is extremely hazardous. Remember

to always wear gloves and dispose of waste in the proper containers

Objectives:

Students will learn to:

Identify all equipment and Chemicals needed for process weigh chemicals to the specific amount needed for process dissolve chemicals to full saturation properly prepare sample properly place gel into the Electrophoresis chamber Properly load the DNA into the gel. properly run the prepared gel through Electrophoresis document results

Self Assessment / Training Assessment: Gel Electrophoresis

The user is able to correctly identify the equipment and Supplies usedo Weigh Boato Analytical balanceo Molecular Weight laddero TAE buffero 8 tooth combo Gel casting trayo Para filmo A 2 to 20 µl pipette and tipso Spatulao Loading Dyeo Microwaveo Voltage adapter Power supply

The user is able to correctly dissolve Gel according to procedureso Measured precisely the proper amount of gelo Dissolved the gel completely using proper procedures

The user is able to prepare Gel for sample according to procedureso Properly sealed gel casting trayo Properly placed 8 tooth comb into trayo Carefully solidified gel casting trayo Filled Horizontal electrophoresis chamber with TAE buffero Removed 8 tooth comb without cracking gelo Correctly placed casting tray into horizontal electrophoresis chamber

The user is able to correctly demonstrate proper loading procedure o Produced viable DNA sample from storageo Prepared Para film for correct use according to procedureo Mixed proper amount of DNA with loading dyeo Correctly placed entire sample into wellso Sealed and ran gel sample

The user is able to correctly stain and document their sampleo Able to set up staining station for proper useo Correctly removed the casting tray from chambero Placed gel into stain solution without harming or breaking gelo Stained sample for correct amount of timeo Removed gel from stain bath to de-staining bath without disrupting gelo De-stain gel for 10 minuteso Placed gel into documentation chamber using proper procedureo Adjusted image so gel was clearly seen

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o Documented gel using UV light in order to receive a discernable imageo Properly disposed of gel according to proper disposal procedures

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Gel Electrophoresis Safety Checklist

The user has demonstrated proper lab safety considerations.o Wash hands thoroughly before processo Wearing gloves and safety glasseso Wearing proper dress attire for lab safety

The user gave a general explanation of all safety protocols for;o Transilluminator machineo Biohazard safety and disposal techniqueso Chemical handling and disposal techniques

The user has demonstrated proper Transilluminator safety precautions.o Wearing full UV light masko Covering all exposed skino Wearing nitrile gloves

The user demonstrated proper equipment use during procedure.o Electrophoresis chambero Transilluminator machine

The user has demonstrated proper Chemical handling and Biohazard safety.o Did not remove gloves while touching potential hazardous chemicals or agentso Did not touch face or exposed skin after chemical or biohazard handlingo Cleaned/wiped up spills promptly and thoroughlyo Correctly identified materials containing biohazardso Handled biohazard materials carefully o Handled biohazard materials according to precautions

The user demonstrated proper clean-up and Chemical / Biohazard disposal o Identified chemicals needing special disposalo Disposed of chemicals according to specific chemical handlingo Disposed of biohazard materials according to proper procedureo Properly cleaned station and all glassware usedo Properly stored all equipment and supplies usedo Correctly cleaned and maintained all equipment used

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Trainer Signature__________________________________________Date:_________