molecular biology
studying biology at the molecular level, studies proteins and genes
genes
sections of DNA
second definition of molecular biology
describes the techniques used to understand how genes and proteins work; often involving DNA manipulation
central dogma of molecular biology
describes the normal flow of genetic info
replication
DNA --> DNA
transcription
DNA --> RNA
Translation
RNA --> protein
DNA polymerase
What enzyme is involved in replication?
RNA polymerase
What enzyme is involved in transcription?
occurs at the ribosome
What organelle is involved in translation?
DNA polymerase
copies both strands of DNA
RNA polymerase
copies one strand of DNA --> mRNA (messenger RNA)
mRNA
the code used to make proteins
protein
amino acids joined together in polymers
some viruses can replicate their own DNA, and retroviruses
What are the exceptions to the normal flow of genetic information?
RNA Dependent RNA Polymerase
What is the enzyme involved in viruses replicating their own DNA?
Retrovirus
inserts the virus into the DNA
Reverse Transcriptase
What is the enzyme involved in the exception of a retrovirus?
DNA
a polymer made up of nucleotides
sugar (deoxyribose), phosphate group, and 1 of 4 nitrogeneous bases
3 components of nucleotides
phosphodiester bonds
nucleotides connect on each strand using the phosphate groups
T
A goes with ____ and vice versa
G
C goes with ____ and vice versa
complement, reading the other
The base pairs _______ each other, so you can get one strand by ______________.
recombinant DNA
putting DNA from one organism into a different organism
Have a DNA template (usually as a plasmid)
First Step of Recombinant DNA Technology
Use restriction enzymes to "cut out" section of DNA you want
Second Step of Recombinant DNA Technology
Use a ligase to join a fragment with a new genome
Last Step of Recombinant DNA Technology
restriction enzyme
breaks phosphodiester bonds
fragments
cut out sections
ligase
brings about the binding and joining of two molecules
precise
Unlike other forms of genetic engineering, CRISPR is super ______.
Guide RNA (gRNA), Cas9 enzyme
What are the two key molecules of CRISPR?
guide RNA
100 nucleotide RNA --> last 20 nucleotides match up with target DNA sequene
cas9 enzyme
cuts DNA at the site that matches with the gRNA
Once Cas9 cuts the DNA, the cell recognizes that it's broken and inserts the new DNA fragment at the cut site.
CRISPR process
Gel Electrophoresis
separates DNA fragments by size
pore size and DNA fragments
running buffer
conducts the electrical current; used to make a fill electrophoresis chamber
TAE
mix of tris, acetic acid, and EDTA
DNA size standard
has fragments of known lengths to be able to compare
used to estimate sample lengths
What is the DNA size standard used for?
power supply
controls the amount of electricity going to the electrophoresis chamber; run at a specific voltage or current based on the gel