Gel Electrophoresis Flashcards
How many steps does a Gel Electrophoresis have?
5 Steps
What are the Gel Electrophoresis steps?
1) Make the gel
2) Set up the gel apparatus
3) Load the DNA sample into the gel
4) Hook up the electrical current and run the gel
5) Stain the gel and analyze the results
Are all DNA strands the same length?
No, DNA strands have several different lengths
Are DNA strands able to be seen under microscopes?
No, DNA strands are molecules so tiny that you can’t see them under most microscopes
What even is Gel Electrophoresis?
Sorting and measuring DNA strands tha you can’t see or touch
When do scientists use Gel Electrophoresis?
Whenever DNA strands need to be sorted according to length
What is Gel Electrophoresis useful for?
It’s a useful technique when separating other types of molecules, like protiens
What does the gel do?
The gel in the filter sorts out the DNA strands
What is Electrophoresis?
Electrophoresis is how DNA strands are pushed throught the gel filter, adding electrical current to make DNA move
Do short strands move faster or slower than longer strands in gel?
Short strands move through the holes in the gel quicker than the long strands
Do longer strands move farther away from the starting point or do shorter strands?
Overtime shorter strands will move farther away from the starting point than the longer strands
What happens when strands are the same length?
DNA strands of the same length will move at the same pace and end up being grouped together
How are DNA strands sorted out?
DNA strands sample sort themselves
Can you see DNA?
No, but you can see large groups of stained strands, these groups show up as bands in the gel
What are the things needed to make the gel?
- powdered seaweed agarose
- buffer
- a flask
- a microwave
- gel mold
- gel comb
What are the eight steps in #1) Making the Gel?
1) Put small amount of agarose in flask
2) Add liquid buffer
3) Loosely place plastic wrap over flask (to prevent overflow)
4) Place flask in microwave and heat mixture (till agarose melts into buffer)
5) Remove plastice wrap and pour agarose mixture into mold
6) Place comb into the gel on one end
7) Let gel cool and solidify (usually takes around half an hour) (as gel cools tiny holes will form in it)
8) Carefully remove comb (leaving empty wells for DNA samples)
What is buffer?
Buffer is a salt water solution that will let electrical charges flow through the gel
What are the two steps in #2) Set Up Gel Apparatus?
1) Pour buffer into electrophoresis box
2) Place the gel still in the mold, into box (the gel should barley be submerged in the buffer)
What are the things you will need for step #2? (Set Up Gel Apparatus)
- the gel you just made
- an electrophoresis box
- another bottle of buffer
What does the buffer do in step #2?
The buffer will conduct the electrical current from one end of the gel to the other, this will also keep the gel from drying out the experiment
What are the five steps of #3) Load the DNA sample into the gel?
1) Suck loading buffer with clean pipette and add it to the DNA sample
2) Suck up some of the DNA sample into the pipet tip
3) Eject the DNA sample from the pipet into the first well of the gel
4) Using a clean pipet tip, suck up some of the DNA size standard
5) Transfer DNA size into the next empty well
What are the things needed for step #3) Load DNA sample into the gel?
- loading buffer
- tube of DNA
- DNA size standard
- micropipette
- electrophoresis box containing buffer and gel
- pipette tips
What and why are DNA samples prepared in clear liquid solution?
Because they would be hard to see if you tried to load it directly into a well
What does the dye in the buffer do?
It makes the sample easier to see and also slightly goopy
