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This is a handout for my ppt presentation about gel purification and western blot
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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
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