Quizs- ABE Lab 2a & 4a

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

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

• False; Restriction enzymes are only found in Eukaryotes.

• True; Restriction enzymes are synthetic and not found in nature.

• False; Restriction enzymes are only found in viruses.

Purpose of DNA probe/stain in lab 4a

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

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

• Probe binds to proteins, Allows you to see the protein structures under UV light which are otherwise clear

• Probe binds to lipids, Allows you to see the lipid layers under UV light which are otherwise clear

purpose of DNA Ladder

• A DNA ladder is a molecular weight marker used to identify the approximate size of a molecule run on a gel during electrophoresis.

• A DNA ladder is used to amplify DNA samples for cloning purposes.

• A DNA ladder serves as a template for DNA synthesis in PCR reactions.

• A DNA ladder is a type of enzyme used to cut DNA at specific sequences.

In lab 4a, how will you know there are Different forms of plasmids present including nicked, multimer, supercoiled, and "regular" plasmids?

• The will move at different rates based on their size and/or shape. The multimer is the biggest since it is 2 plasmods connected together; it will move the slowest. The nicked, regular, and supercoiled are all the same size but will travel at different rates based on their shape. The supercoiled acts linear so it moves the fastest.

• The multimer is the biggest and will move the slowest.

• The nicked plasmid is the fastest moving form.

• The supercoiled plasmid moves slower than the regular plasmid.

In lab 4a, what do you expect to see in the R- lane and why?

• Only full plasmids, possibly as separate bands or a smear due to different plasmid configurations.

• Only linear DNA fragments due to restriction enzyme digestion.

• A mixture of RNA and DNA bands indicating contamination.

• No bands at all, indicating a failed experiment.

How many Restriction fragments will there be after digest with Hind III and BamH I

• 2 fragments

• 2 fragments

• 3 fragments

• 4 fragments

In lab 2a, what was similar and different between the R+ and R- tubes?

• Both R+ and R- tubes had the pARA-R plasmids in them, and the R+ tube had restriction enzymes in it while the R- tube did not.

• Both R+ and R- tubes had different plasmids in them, and the R+ tube had restriction enzymes in it while the R- tube did not.

• Both R+ and R- tubes had the same plasmids in them, and the R- tube had restriction enzymes in it while the R+ tube did not.

• Both R+ and R- tubes had the pARA-R plasmids in them, and the R- tube had no enzymes while the R+ tube had enzymes.

Describe the parts of the pARA-R plasmid

• The pARA-R plasmid is 5,302 bp long and includes the green fluorescent protein (gfp) gene, the kanamycin resistance gene (kanR), and the arabinose activator (araC).

• 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).

• The pARA-R plasmid is 4,500 bp long and includes the red fluorescent protein (rfp) gene, the ampicillin resistance gene (ampR), and the lactose operon (lacO).

• The pARA-R plasmid is 5,302 bp long and includes the blue fluorescent protein (bfp) gene, the chloramphenicol resistance gene (cat), and the arabinose activator (araC).

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

• HINDIII and BamHI

• EcoRI and NotI

• XhoI and KpnI

• SmaI and SacI

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

• To ensure the plasmid is correct for making the red fluorescent protein by using restriction enzymes and verifying fragment sizes.

• To analyze the DNA sequence of the plasmid without any modifications.

• To amplify the plasmid DNA for further experiments without any verification.

• To clone the plasmid into a bacterial host for protein expression.