How to Make an Electrophoresis Gel
Find the gel concentration required., Obtain an electrophoresis gel casting tray., Gather the required chemicals., Add the agarose., Prepare the mixture., Add the EtBr., Fill the casting trays., Insert the combs., Allow the casting trays to cool and...
Step-by-Step Guide
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Step 1: Find the gel concentration required.
This must be specified by the physician and will vary based on the type of testing required by the study the physician is performing.
Common concentrations are
0.7 percent.
0.8 percent,
1.0 percent and
1.2 percent. -
Step 2: Obtain an electrophoresis gel casting tray.
These trays are available at medical supply outlets.
Note the volume of the tray because it will determine the maximum amount of electrophoresis gel that can be made at once. , Get an amount of Tris-Acetate-EDTA (TAE) sufficient to make up the volume of the gel being made.
The amount of agarose, the polymer that forms the gel, will depend on the concentration of the gel to be made.
The expressed gel percentage is agarose in grams divided by TAE in mL.
The final ingredient needed is ethidium bromide (EtBr).
EtBr makes DNA visible under ultraviolet light.
Divide the amount of TAE used by 1000 to determine the amount of EtBr needed. , Add the determined amount of agarose to the required amount of TAE.
Gently stir or swirl the mixture to ensure proper mixing. , The agarose in TAE must be heated in a microwave oven to dissolve it.
Warm the solution for short periods of time, such as 15 seconds, on low power.
Check the progress of the melting constantly.
If the TAE is warmed to the point where particulate matter is formed, the batch has been ruined. , EtBr is a powerful biohazard.
Laboratory gloves should be worn for this part of the process.
Add the EtBr after heating of the TAE has been completed.
Gently stir or swirl the container to ensure proper mixing.
Allow the liquid to stand and cool for 5 minutes. , Swirl or stir the chemical mixture to ensure that there is uniform temperature throughout the mixture.
Pour the mixture carefully in to the casting tray. , Place the casting combs into the support rails of the casting tray.
The comb teeth will create wells in the gel to which the sample to be tested can be added.
The combs come in various sizes to create various size wells.
The size wells that need to be formed will be determined by the physician based on the study to be performed. , Remove the casting combs.
Remove the gel from the casting tray.
The electrophoresis gel is now ready for use. -
Step 3: Gather the required chemicals.
-
Step 4: Add the agarose.
-
Step 5: Prepare the mixture.
-
Step 6: Add the EtBr.
-
Step 7: Fill the casting trays.
-
Step 8: Insert the combs.
-
Step 9: Allow the casting trays to cool and the gel to set for 1 hour.
Detailed Guide
This must be specified by the physician and will vary based on the type of testing required by the study the physician is performing.
Common concentrations are
0.7 percent.
0.8 percent,
1.0 percent and
1.2 percent.
These trays are available at medical supply outlets.
Note the volume of the tray because it will determine the maximum amount of electrophoresis gel that can be made at once. , Get an amount of Tris-Acetate-EDTA (TAE) sufficient to make up the volume of the gel being made.
The amount of agarose, the polymer that forms the gel, will depend on the concentration of the gel to be made.
The expressed gel percentage is agarose in grams divided by TAE in mL.
The final ingredient needed is ethidium bromide (EtBr).
EtBr makes DNA visible under ultraviolet light.
Divide the amount of TAE used by 1000 to determine the amount of EtBr needed. , Add the determined amount of agarose to the required amount of TAE.
Gently stir or swirl the mixture to ensure proper mixing. , The agarose in TAE must be heated in a microwave oven to dissolve it.
Warm the solution for short periods of time, such as 15 seconds, on low power.
Check the progress of the melting constantly.
If the TAE is warmed to the point where particulate matter is formed, the batch has been ruined. , EtBr is a powerful biohazard.
Laboratory gloves should be worn for this part of the process.
Add the EtBr after heating of the TAE has been completed.
Gently stir or swirl the container to ensure proper mixing.
Allow the liquid to stand and cool for 5 minutes. , Swirl or stir the chemical mixture to ensure that there is uniform temperature throughout the mixture.
Pour the mixture carefully in to the casting tray. , Place the casting combs into the support rails of the casting tray.
The comb teeth will create wells in the gel to which the sample to be tested can be added.
The combs come in various sizes to create various size wells.
The size wells that need to be formed will be determined by the physician based on the study to be performed. , Remove the casting combs.
Remove the gel from the casting tray.
The electrophoresis gel is now ready for use.
About the Author
Sara Moore
Writer and educator with a focus on practical crafts knowledge.
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