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Lab Report Concept Map

Grow and Copy

Cell Culture

Dilutions

Spinning Down

Spin down in centrifuge

Remove supernatant

Resuspend cell pellet

Less concentrated volume from high concentrated volume

Lag Phase

Cell acclimation

Log Phase

Increase in cells

Stationary Phase

Nutrients depleted

Growth slows

Death Phase

Deadly conditions

Cell death

OD reading

Concentration of cells in culture

Plasmids

Circular DNA

Separate from Chromosomal DNA

Environmental Pressure

Lost

Kept

Aseptic Technique

Prevents Contamination

Spotting Assay

Growth Rate Test

Growth Conditions

Different Media

3 Way Streaking Technique

T-streak

Selectable Marker

Yeast

Foreign DNA uptake

Caused by stress

In chromosomes

Take place of genes

Homologous Recombination

Bacteria

Gene with phenotype that can be selected for

Usually on plasmids

Transformation

Heat

Electricity

Chemical Treatment

PCR

Synthesize DNA

Isolate and amplify

Steps

Extension

Polymerase

Copy template

Double stranded DNA created

Annealing

polymerase binds

primer/DNA template

Primer binds ssDNA

Denaturation

Denature primers

Denature DNA

DNA splits

ssDNA

Initialization

Activate Polymerases

denature DNA template

Ingredients

Buffer

Maintain pH

DNA denaturing

DNA renaturing

Polymerase activity

dNTPs

DNA building blocks

Create DNA copies

Primer

Single Strand

Binds template DNA

Reverse

reverse complement of template

endpoint for PCR

Forward

Matches template

Starting Point PCR

DNA Template

Read by polymerase

cDNA template

Plasmid template

Genomic Template

DNA Polymerase

New DNA strand

From template

PFU

Taq

Thermocycler

Machine

Temperature changes

Annealing Temperature

primers pair with DNA

Melting Temperature

primers dissocitate from template

Histones

Variants

LUCA

FECA

LECA

Urkaryote

Cannonical

H2A

H2B

H3

H4

H1

Reagents

Multiple Chemical Solutions

Calculate stock amounts

Calculate solvent amount

Add solvent and chemicals

Molar/percent solutions (liquid)

Subtract stock from total volume

Add exact stock

Add exact solvent

Percent solutions (powder)

Weight of Water

w/v

w/w

Add weight

Dissolve

Final Volume

Liquid Stocks

v/v

Molar solutions (powder)

Concentration Mol/L

Dissolve Powder

Adjust pH

Exact Volume

Visualize DNA

Gel and Electrophoresis

Characterize DNA and RNA

TAE Tris, Acetic Acid, EDTA

Maintain pH levels

Agarose Powder + Buffer

Microwave Mixture

Add Ethidium Bromide

Light Fluorescence

Intercalate between DNA

Cast Gel

Dye DNA

Load DNA

Charge applied

DNA moves to positive charge

Strands separate

Strands separate based on size

Short strands are faster

DNA Ladders

DNA size by comparison

100 bp

1 kb

DNA and RNA Extraction

Ethanol precipitation

Desalt and Concentrate

Isolate

DNA

Nanodrop

Purity

Concentration

Phenol Chloroform

Denature proteins

Extractable DNA

Slightly alkaline

RNA

Uses acidic

Cell Lysis

Mechanical

Douncing

Sonication

Bead Beating

Yeast Cell Lysis Buffer

Zymolyase

Break yeast cells

Tris

No pH Change

EDTA

Chelator

prevent enzymatic function

SDS

Detergent