Lab Quiz 3 Flashcards
Lab 3 and 4
What is the buffer in electrophoresis?
It is a buffer of ions that allows for the conduction of current.
What is a palindrome?
What is the significance of it?
It is a sequence that is read backwards when the strands are in opposite directions.
‘5-AGTACT
TCATGA-5’
Restriction enzymes cut at certain palindromes
What is agarose gel composed off?
It is seaweed extract which is a polysaccharide
What is the loading dye responsible for?
It provides density so that our sample will sink into the wells, as well as giving us visual cues to the progression of the electrophoresis.
What is an intercalating agent?
It is an agent that will stick between the bases of the DNA and when it is exposed to UV light will get excited and fluores
What is ethidium bromide?
Why is it more harmful than something like SYBR?
It it an intercalating agent that is a known mutagen and carcinogen.
It is harmful because it can readily pass biological membranes and cause mutations.
What is SYBR?
It is the new intercalating agent that is noncarcinogenic - it doesn’t have the ability to get into the membrane.
How much of our sample did we use per test tube?
We used 100ng of our vector per tube.
What was the ladder we used to compare our samples against?
500, 1000, 1500, 2000, 2500, 3000, 4000, 5000, 6000, 8000, 9000, 10k
What was the enzyme that we used in the original cutting?
We used BAM H1
If a particular vector map does not show where a specific enzyme cuts what does this mean?
It means that the enzyme cuts the vector in many places.
What is the purpose of running the uncut DNA?
It provides a comparison from the supercoiled DNA to that of the linear, cut DNA.
It allows us to confirm that the digestion actually worked.
