exam 2 biol 107 csun

Oglionucleotides

Small DNA fragments

dNTPs

free base pairs that serve as building blocks for DNA synthesis and replication

Taq polymerase enzyme

synthesizes new DNA strands from DNA template, produces complementary strand

Buffer (no enzyme)

maintains optimal pH and ionic conditions for enzyme activity, ensuring efficient DNA amplification during the reaction

PCR

Polymerase chain reaction

PCR step 1 (name, temp degrees C)

Denaturation, 94 degrees C

PCR step 2 (name, temp degrees C)

Annealing, 55 degrees C

PCR step 3 (name, temp degrees C)

Extension, 72 degrees C

Denaturation

DNA strand separates into two single strands

Annealing

DNA primers attach to DNA template

Extension

Taq polymerase enzyme make new strand DNA

Gel electrophoresis

Used to separate DNA fragments based on size

Small molecules

Move faster and travel farther through gel

ADP-1

wild type, no mutation

ADP-6

mutated, lacks enzyme to metabolize pHB

Succinate

Carbon source both ADP strains can grow on

Natural transformation

naturally occuring, bacterium will pick up foreign DNA and incorporate it into its genome

Unatural transformation

Heat shock and electroporation

Transformation: Translocasome

in cell envelope, upatakes DNA

Transformation: induce competence

Secretes signal, competence factor, stress (starvation)

Transduction

Bacteriphage injects DNA into cell, either viral DNA or bacterial DNA accidentally

Conjugation

Bacterial sex, bacteria transfer via sex pilus, pilus proteins encoded on F factor, F- female acceptor becomes F+ male donor

Plasmid

Genetic structure in a cell that can replicate independently of the chromosome

Plasmid: origin of replication

Necessary for plasmid copying during bacterial replication

Multiple Cloning Site (MCS)

location for inserting gene of interest

Sleectsble marker

Gene encoding a protein that will make the bacteria resistant to an antibiotic or allow growth of the bacteria in the absnese of food source

Plasmid IRL use

Mini factories to make many copies of DNA

Green fluorescent protein (GFP)

Originaly from hellyfish, encoded on plasmids with ampilicilin resistance

Beta lactamase

Enzyme that makes bacteria ampicillin resistant, destroys ampicillin structure

Ampicillin

Prevents the synthesis of cell wall

Lab 6 transformation steps

Isolate DNA from wild type, dilute DNA, mix with mutant bacteria

Lysis buffer

Breaks open cell, cloudy

3 ways bacteria exchange genetic information

Transformation, transduction, and conjugation

Codon

3 nucleotides

4 nucleotides of DNA

A-T

G-C

Is DNA antiparallel

Yes