Araullo University-Phinma Education Network

College of Education

Gel Purification and

Western Blot

Prepared by:

Jan del Rosario

3SED-SC

Submitted to:

Mrs. Lucy Rivera

Gel Purification

-also known as gel extraction or gel isolation

-is a technique used to isolate a desired fragment of intact DNA from an agarose gel.

4 Basic Steps

1. identifying the fragments of interest 2. isolating the corresponding bands 3. isolating the DNA from those bands 4. removing the accompanying salts and stain

How It’s Done

1. UV light is shone on the gel in order to illuminate all the ethidium bromide-stained DNA.

2. The desired band is identified and physically removed with a cover slip or razor blade.

3. The removed slice of gel should contain the desired DNA inside. 4. isolating and cleaning the DNA fragment of interest

Techniques in Isolating the DNA Fragment

Spin Column Extraction Dialysis Traditional

Significance

Cloning Production of recombinant DNA Genetic Engineering

Western Blot

- also known as protein immunoblot

- is a widely used analytical technique used to detect specific proteins in a sample of tissue homogenate or extract.

Steps

1. Tissue preparation 2. Gel electrophoresis 3. Transfer 4. Blocking 5. Detection 6. Analysis

Uses

HIV Test or Lyme disease in blood samples Epitope mapping, amino acid composition and sequence analysis