Flashcards- ABE Lab 2a & 4a

Front

Overall Purpose of LAB 2a (restriction digest) and LAB 4a (gel electrophoresis)

Back

Lab 2a and 4a are needed to ensure the plasmid is correct for making the red fluorescent protein. Lab 2a uses restriction enzymes to cut the plasmid, and lab 4a is used to verify that these are the correct size fragments for the para-R plasmid.

Front

Use of restriction enzymes in LAB 2a

Back

A combination of 2 different restriction enzymes is used to digest (hydrolyze) the pBAD-rfp segment out of the plasmid.

Front

Describe the parts of the pARA-R plasmid

Back

The pARA-R plasmid is 5,302 bp long and includes components like the red fluorescent protein (rfp) gene, the ampicillin resistance gene (ampR), and the arabinose activator (araC).

Front

True or False: Restriction enzymes are naturally found in Prokaryotes.

Back

True; Restriction enzymes provide protection against invading viruses by hydrolyzing "foreign" DNA.

Front

Restriction enzymes cut DNA at specific recognition nucleotide sequences called ________

Back

restriction sites

Front

What are "sticky ends"

Back

Restriction enzymes like BamHI, HindIII, and EcoRI cut DNA at specific sequences, resulting in "sticky ends" which are single stranded overhangs that can be used for DNA recombination.

Front

Which 2 Restriction enzymes did we use in lab 2a to digest the pARA-R plasmid?.

Back

HINDIII and BamHI

Front

Purpose of LAB 4a

Back

Use gel electrophoresis to separate out the DNA fragments; Examine the products from the restriction digest of the pARA-R plasmid

Front

Purpose of DNA probe/stain in lab 4a

Back

Probe binds to DNA, Allows you to see the DNA fragments under UV light which are otherwise clear

Front

Purpose of loading dye in Lab 4a

Back

Loading dye adds weight to the DNA which helps it sink into the well, it also allows you to track general movement in the gel box (it does not bind to the DNA so you cannot actual see the DNA moving, you can only infer that it is)