Molecular Biology - Unit 1

Topics 1-7

What are key structures of features of DNA & RNA nucleotides

Triphosphate group, sugar group and a base

Bases in DNA

Adenine, Thymine, Cytosine, Guanine

Sugar in DNA

Deoxyribose

Phosphodiester bonds

Covalent bonds connecting individual nucleotides

How are base pairs formed

Non covalent hydrogen bonds form base pairs to connect two individual polymers

Bases in RNA

Adenine, Uracil, Cytosine and Guanine

Sugar in RNA

Ribose

What groups make a protein

C surrounded by an amino group, hydrogen, carbonyl gorup and R gropu

Primary structure

polymer of amino acids, connected by peptide bonds

Secondary Structure

Hydrogen bonds between R groups

Tertiary Structure

3D shape of a protein, mostly driven by hydrogen bonds and other non covalent interactions

Quaternary Structure

Two or more proteins interacting “permanently”

Three steps of DNA extraction

Lysis, separate DNA out of all the cell junk, washing and eluting the DNA

Lysis

Getting DNA out of cell and out of nucleus

Chaotropic Salts

Disrupt the noncovalent interactions in membranes

Proteases

Enzyme that digests protein

Silica in column

Silica resin-DNA sticks, everything other than DNA flows out of column

Chelex Beads

DNA stays in solution fo water, chelex beads, everything other than DNA sticks to beads

Denaturation

Temperature is increased to separate DNA strands

Annealing

Temperature is decreased to allow primers to base pair to complementary DNA template

Extension

Polymerase extends primer to form nascent DNA strand

Exponential Amplification

Process is repeated and region of interst is amplified exponentially

Reaction ingredients

Template DNA, Polymerase Enzyme, Primers, Free Nucleotides

Electrophoresis

the movement of charged particles in a fluid or gel under the influence of an electric field.

Longer slower molecules in gel

near top

Shorter faster molecules in gel

near bottom

GMO

geneticallly modified organism

RTPCR

Reverse transcriptase PCR

Quanitative PCR

How much of a certain sequence is present

Endpoint/Conventional PCR

Is a certain sequence present

Gene Expression in PCR

requires process of transcription which involves RNA (mRNA)

EMSA

Electrostability Shift Assay

Western Blotting

To visualize specific proteins on the gel