Lab 6
linear
A plasmid state that travels according to it's true size
relaxed
A plasmid state where the molecules are a large floppy circle that travels slowly through the gel.
supercoiled
A plasmid state where the molecules are twisted up very tightly like a rubber band and travels quickly through the gel.
heterozygous
Are more people in the population homozygous or heterozygous?
negative, positive, smaller
DNA, because it has a ___ charge, moves to the ___ end (anode) of the field in gel electrophoresis. ___ DNA molecules migrate most quickly
positive, anode, sugar-phosphate backbone
During gel electrophoresis, DNA fragments move towards the ___ electrode, also called the ___ because of the negative charged carried by their ___ ___ ___
bands
During gel electrophoresis, fragments of different sizes will separate, forming distinct ___ on the gel
2
If an individual is heterozygous how many bands will appear in the lane?
1
If an individual is homozygous how many bands will appear in the lane?
nontransfromed cells grew together and form a lawn
Individual colonies are not visible on you LB only plate. Instead a white smear is visible. This is expected because...
allow you to see how fast your DNA is migrating through the gel
Purpose of dyes
flooded with water
Removing the comb from a gel is made much easier, with less possibility of tearing the gel, if the area around the teeth of the comb is...
single digest
Term to describe using one restriction enzyme on a plasmid
double digest
Term to describe using two restriction enzymes on a plasmid
slower
The larger the DNA fragments, the ___ it will move through the gel
supercoiled, relaxed, linear
The speed of a circular plasmid (one that has not been cut by a restriction enzyme) will depend on whether the plasmid is...
hydrolyzed
The term "digested" when talking about restriction enzymes actually means what?
EcoRI and EcoRV
The two restriction enzymes in Lab 6
run to red
The well containing DNA samples are placed far from the anode and can "___ ___ ___"
15-20
To melt solidified agarose in a bottle in the microwave, heat it in short intervals of ____ seconds each
DNA ladder
Tool to determine the actual length of the DNA in number of base pairs.
inserting the comb incorrectly at the adjustable end
Upon removal, what can cause the cast to break at the wells and become unusable?
digested
What happens to foreign DNA entering the bacterial cell once the restriction site is detected by the restriction enzyme?
dye and reagent glycerol
What is in the loading dye?
16(r) + 146
What is the equation for total allele size?
makes sample denser than the runner buffer
What is the purpose of the reagent glycerol in the loading dye?
primer dimers, bottom, small, quickly
What primers, instead of annealing to target DNA sequence, annealed to one another? They are found at the ___ of the gel because they are ___ and travel very ___ through gel.
gel electrophoresis
What process physically separates molecules from one another based on structural features such as seize and charge?
small, to slow the fragments down
What size fragments of DNA do you want thick gel and why?
large, to allow fragments to migrate through the gel
What size fragments of DNA do you want thin gel and why?
3000, 1000, 500
What three bands did you select on the DNA ladder?
during PCR
When are primer-dimers produced?
use a sharpie to place a dot over each colony as you count it
When counting the colonies on your bacterial transformation plates, the best way to keep track of which ones have been counted is on the backside of the plate is to...
loosened
When microwaving a concealed solution of agarose should the cap on the bottle be tightly closed or slightly loosened?
