PCR
What does PCR stand for?
Polymerase chain reaction
Name the components of a "regular" gel from gel electrophoresis.
Agarose and 1X TAE.
What are the monomers of proteins?
Amino acids.
What type of instrument is used to complete a PCR amplification?
Thermocycler.
In gel electrophoresis, what does the gel act like?
A filter/sponge/sieve.
Lipids are composed of which elements?
Carbon, hydrogen, and oxygen.
Why can’t we use human DNA polymerase for PCR?
Human polymerase cannot withstand the temperatures needed for PCR to work.
In gel electrophoresis, how do we make the DNA migrate through the gel?
We apply an electrical current, i.e., a negative charge near the wells ad a positive charge away from the gels.
What do you are call an extragenomic piece of DNA commonly found in bacteria?
A plasmid.
Why do you need two primers for PCR?
The primers need to flank the target region (i.e., region that should be copied). Otherwise, the Taq polymerase will copy more DNA than necessary and/or miss the target region entirely.
What is the purpose of the DNA ladder during gel electrophoresis?
To help scientists estimate the size of the bands from their samples.
Which level of protein structure remains even after a protein has been denatured?
Primary, i.e., the amino acid sequence.
Why do you need to do PCR before doing gel electrophoresis?
A lot of DNA is needed to form visible bands on the gel.
What is the purpose of using restriction enzymes before doing gel electrophoresis?
It generates fragments of DNA that can be analyzed using the gel.
Why is DNA replication considered semiconserative?
Because one strand of DNA is used as a template for the new strand.
Name the stages of PCR and their associated temperatures in order.
(1) Denature - 94-98
(2) Anneal - 50-65
(3) Extend - 72-80
If we have a baby, and we know who the mother is but we want to determine the father using gel electrophoresis, then how will we know we have found the father?
The father will match the baby at any band where the baby does not match the mother.
In eukaryotes, what happens to mRNA after it's been transcribed?
RNA processing (introns removed; poly A tail and 5' guanosine cap added).
Name the five components in the PCR reaction mix.
Taq polymerase, dNTPs, primers, buffer, sample
Name three differences between "regular" gel electrophoresis and PAGE.
The gels from "regular" gel electrophoresis are made of agarose, are used to separate DNA molecules only, is run horizontally, and have a lower resolution compared to PAGE.
The gels in PAGE are made of polyacrylamide, can be used to separate out proteins (if SDS is used) as well as DNA molecules, is run vertically, and has a higher resolution than "regular" gel electrophoresis.
What happens when a repressor binds to its operator?
Transcription stops.