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PCR and Electrophoresis Handout
PCR and Electrophoresis Handout
PCR and Electrophoresis Handout
• Basic Steps:
1. _________________________– to expose the DNA within
2. _________________________ – salting out procedure
3. _________________________ – using PCR
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• Buffer • Gels
• To carry the current and • Agarose gel
______________ during electrophoresis • Polysaccharide polymer extracted from
•A solution of seaweed
_________________________________
_________________________________ • Polyacrylamide gel
• pH is constant as it takes up or release • Acrylamide in combination with cross-
protons linker methylene bisacrylamide
• Tris buffers: Tris borate EDTA, Tris • Best for very small DNA fragments and
single stranded DNA
phosphate EDTA, Tris acetate EDTA
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• The charge on the molecules to be • Other Separation technique for DNA and RNA detection
separated leads them to move • Radioactive labelling
through the gel toward an electrode
of the opposite charge.
• Autoradiography
• The separation are based on the size
and the sieving action of the gel.
• Chemiluminiscent
• The Smaller oligonucleotide the
farther it moves than the larger one.
• Fluorescence
PCR
• Developed in 1983 by Kary B. Mullis
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