Lab Report Concept Map

DNA and RNA Extraction

Cell Lysis

Yeast Cell Lysis Buffer

SDS

Detergent

EDTA

Chelator

prevent enzymatic function

Tris

No pH Change

Zymolyase

Break yeast cells

Mechanical

Bead Beating

Sonication

Douncing

Phenol Chloroform

Denature proteins

RNA

Uses acidic

Extractable DNA

Slightly alkaline

Nanodrop

Concentration

Purity

Ethanol precipitation

Desalt and Concentrate

Isolate

DNA

RNA

Gel and Electrophoresis

DNA Ladders

DNA size by comparison

1 kb

100 bp

Agarose Powder + Buffer

Microwave Mixture

Add Ethidium Bromide

Intercalate between DNA

Cast Gel

Dye DNA

Load DNA

Charge applied

DNA moves to positive charge

Strands separate

Short strands are faster

Strands separate based on size

Light Fluorescence

TAE Tris, Acetic Acid, EDTA

Maintain pH levels

Characterize DNA and RNA

Molar solutions (powder)

Concentration Mol/L

Dissolve Powder

Adjust pH

Exact Volume

Percent solutions (powder)

Liquid Stocks

v/v

Weight of Water

w/w

Add weight

Dissolve

Adjust pH

Final Volume

w/v

Add weight

Dissolve

Adjust pH

Final Volume

Molar/percent solutions (liquid)

Subtract stock from total volume

Add exact solvent

Add exact stock

Multiple Chemical Solutions

Calculate stock amounts

Calculate solvent amount

Add solvent and chemicals

Cannonical

H1

H4

H3

H2B

H2A

Variants

LUCA

FECA

LECA

Urkaryote

PCR

Thermocycler

Machine

Temperature changes

Melting Temperature

primers dissocitate from template

Annealing Temperature

primers pair with DNA

Ingredients

DNA Polymerase

New DNA strand

From template

Taq

PFU

DNA Template

Read by polymerase

Genomic Template

Plasmid template

cDNA template

Primer

Single Strand

Binds template DNA

Forward

Starting Point PCR

Matches template

Reverse

endpoint for PCR

reverse complement of template

dNTPs

DNA building blocks

Create DNA copies

Buffer

Polymerase activity

DNA renaturing

DNA denaturing

Maintain pH

Steps

Initialization

Activate Polymerases

denature DNA template

Denaturation

Denature DNA

DNA splits

ssDNA

Denature primers

Annealing

Primer binds ssDNA

polymerase binds

primer/DNA template

Extension

Polymerase

Copy template

Double stranded DNA created

Synthesize DNA

Isolate and amplify

Transformation

Chemical Treatment

Electricity

Heat

Cell Culture

Selectable Marker

Bacteria

Gene with phenotype that can be selected for

Usually on plasmids

Yeast

In chromosomes

Take place of genes

Transformation

Homologous Recombination

Transformation

Foreign DNA uptake

Caused by stress

3 Way Streaking Technique

T-streak

Spotting Assay

Growth Rate Test

Bacteria

Different Media

Growth Conditions

Yeast

Different Media

Growth Conditions

Aseptic Technique

Prevents Contamination

Plasmids

Environmental Pressure

Kept

Lost

Separate from Chromosomal DNA

Circular DNA

OD reading

Concentration of cells in culture

Lag Phase

Cell acclimation

Log Phase

Increase in cells

Stationary Phase

Nutrients depleted

Growth slows

Death Phase

Deadly conditions

Cell death

Dilutions

Less concentrated volume from high concentrated volume

Spinning Down

Spin down in centrifuge

Remove supernatant

Resuspend cell pellet