DNA Structure and Analysis

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