Molecular Biology Techniques: PCR and Restriction Enzymes

Flashcards based on molecular biology techniques, focusing on the processes of PCR, restriction enzymes, and plasmid preparation.

The purpose of SDS detergent is to ___.

disrupt cell membrane and denature proteins.

EDTA in the lysis buffer is used to ___.

remove Mg^2+, destabilize the cell wall, and inhibit DNAses.

NaOH increases pH to ___ and denatures DNA by breaking H-bonds.

~12

During the neutralization step, a ___ solution is used to facilitate the separated gDNA from plasmid.

high salt concentration

Restriction enzymes recognize and cut specific ___ sequences.

palindromic

If there is a single base pair difference in the restriction enzyme site, the enzyme will ___.

not recognize the sequence and will not cut.

In the post-lab question, it is important to select a ___ from the LBAmp plate.

single colony

The gDNA aggregates because it ___ and becomes a solid pellet.

does not renature properly.

BglII is used because it has a cut site on ___ of the insert.

both sides

The reason for not using the entire plasmid prep for restriction digest is to ___.

limit contamination.

The DNA binds to the silica column due to a ___ bridge formed with high salt concentration.

cation

The purpose of doing the restriction digest before sequencing is to ___.

verify the DNA and confirm what is being sent.

BglII cannot be heat inactivated because ___.

it would interfere with sequencing.

The elution solution is used to ___ off the DNA from the silica column.

elute