By: Alex Benjamin Bruce Gel Electrophoresis By: Alex Benjamin Bruce

Restriction Enzyme Restriction endonuclease (Restriction enzyme) is an enzyme that cuts DNA at a specific sequence (restriction site). RFLPs (restriction fragment length polymorphisms) are DNA fragments of different lengths generated by restriction enzyme from the same region of the genome.

Agarose Gel Electrophoresis Put the samples in the well and apply an electric current. Stain the gel with a dye that binds to DNA. The dye fluoresces under UV light.

Mechanism DNA has negative charged backbone. DNA fragments are separated into two strands using agarose gel electrophoresis. Strands move to the positive pole with different speed. The strands are not visible until they are put under UV light.

Factors that affect the speed: Molecular mass (Most important) Current (Voltage) Gel density DNA shape (coil, super coil) Dye Buffer

Analyze Results to map P Male: both homozygous Female: both heterozygous F1: Group 1: heterozygous Group 2: homozygous Group 3: recombinant(1,2,3) Group 4: recombinant(1,2,4) 30 mu between two loci

Applications

Testing proteinuria Proteinuria is the existence of too much protein in urine. The lesion in this kind of cases can be found in glomerulus （if high molecular weight proteins existed) or its mesenchyme (if low molecular weight proteins was found)

Blood test Electrophoresis time shorter than normal Low blood viscosity Tumor or scorbutic Electrophoresis time longer than normal high blood viscosity Thrombus or high blood pressure

Glycosylated hemoglobin Glycosylated hemoglobin is a combination of blood sugar and hemoglobin It was chemically stable, therefore the level of this can represent the long term blood sugar level

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