DNA Extraction

DNA EXTRACTION
LAB SESSION

DNA EXTRACTION

DNA EXTRACTION
 Digestion:

(mechanical, physical, chemical)  Precipitation: Ice cold ethanol  Isolation: TE buffer

GEL ELECTROPHORESIS OF DNA


Separation procedure: electrical field
 Stationary

phase: Agarose Gel (seaweed)  Mobile phase: TBE buffer.


Separation based on:
 Molecule

size  Molecule charge


Migration of DNA from Cathode to Anode.

SEAWEED

CATHODE & ANODE
Cathode: Negatively charged Electrode (or Element) that attracts the Positive (+) ions (Cations) in a Circuit or Chemical reaction. Anode: Positively charged Electrode (or Element) that attracts the Negative (-) ions (Anions) in a Circuit or Chemical reaction. What is the charge DNA? (negatively charged)

GEL ELECTROPHORESIS OF DNA


Agarose gel: Sugar
Cool (polymerization)

Heat (disolve) Polymers crosslink

(Net)  Loading Dye:
Prevent

sample floating Sample monitoring

Ethidium Bromide stains DNA and emit (release) fluorescence if exposed to UV light.  Ladder standard is Known Base Pair (bp.) size used to compare the test product (DNA).


Polymers crosslink (Net)

POLYMERIZTION

POLYMERS CROSSLINKING (AGAROSE MATRIX)

Wells Sample:
1&6 Molecular ladder (Standard) 3. DNA Extract 2&4 DNA PCR product 5. water (negative control)