DNA, RNA, and Proteins!
What is the difference between RNA and DNA?
RNA is single-stranded and DNA is double-stranded.
Also acceptable, U/T
What property of DNA does agarose gel electrophoresis use to separate DNA?
Size!
What is a phenotype?
A trait that is dictated by the environment and one's genotype
Why do we grow bacteria in LB broth?
We grow bacteria in LB broth because it provides a nutrient-rich environment containing peptides, amino acids, vitamins, and salts, which support rapid bacterial growth and replication, especially for common lab strains like E. coli.
What does GFP stand for?
Green fluorescent protein
What is a gene?
A gene is a segment of our DNA that contains information to produce a protein?
What does PCR stand for? Why do we use it?
Polymerase Chain Reaction, to make more copies of DNA!
What do people have different phenotypes?
Because of genetic variation (mutations, varying alleles, etc)
Why do we shake E. coli at 37 C to grow it?
Shaking provides oxygen and 37 C is the ideal temperature for E. coli growth
What is bacterial cell lysis?
A technique to break open cells and release their internal contents
What are the 4 base pairs in DNA?
A, C, T, G
What do restriction enzymes do?
They cut up DNA at specific sequences.
T or F. Sickle cells in sickle cell anemia are considered a phenotype. Explain.
True. While it is not visual right away, it is still a perceivable trait that arises due to the environment and the genotype of an individual
What is the purpose of the ampicillin-resistance gene found in our plasmid that we added to E. coli
To make sure that cells only grow if they contain the plasmid of interest.
In experiment 2, why did we use SDS-PAGE?
To separate proteins from our chromotography results so that we could examine them and see if we successfully obtained GFP!
Are plasmids DNA, RNA, or protein?
DNA
In agarose gel electrophoresis, will DNA migrate to the positive or negative electrode? Why?
In agarose gel electrophoresis, DNA migrates to the positive electrode because DNA has a negatively charged phosphate backbone, which is attracted to the positive charge.
Person A is homozygous dominant for one form of a gene and produces 3 bands in gel electrophoresis. Person B is homozygous recessive and produces 2 bands. What is the maximum amount of bands a heterozygous individual can have?
5 bands
With the exception of overlapping bands if the size is the same
Before growing bacteria on LB- plates, we diluted it heavily. Why?
Since there is no antibiotic in LB-, many bacteria will grow, and we wouldn't be able to see individual colonies
After we lysed our cells, why was our pellet no longer green, but rather our supernatant was green?
Because by lysing the cells we extracted the fluorescent protein into the supernatant
What is the genetic code and what is a codon?
The genetic code is the set of rules by which the sequence of nucleotides in mRNA is translated into the sequence of amino acids in a protein. Each group of three nucleotides (a codon) corresponds to a specific amino acid or a stop signal during translation.
What are the 3 main steps of PCR?
Denaturation, annealing, and extension.
Can be explained in different words
I only have expired PTC strips, but I have access to a molecular biology lab. What are the main steps I need to take to determine whether I'm a bitterness taster? (6 main steps, 5 correct necessary)
Extract cheek cells
Break open/Extract DNA
PCR amplify TAS2R38 gene
Cut TAS2R38 gene with restriction enzymes
Analyze on gel
What is the name of the sterilization machine we used in the basement?
Autoclave
Why do we denature proteins with SDS in SDS-PAGE?
We denature proteins with SDS in SDS-PAGE to unfold them into linear chains and coat them with uniform negative charges, so they can be separated based solely on size (molecular weight) during electrophoresis, rather than shape or native charge.