Gel Electrophoresis and Probes (Southern Blotting) Group A,

321 Before beginning Gel Electrophoresis, test tubes containing identical DNA fragments must be acquired. Test Tubes Restriction Enzyme 1 Restriction Enzyme 2 Restriction Enzymes 1 & 2 Restriction enzymes will be introduced in order to cleave the DNA into fragments of different sizes.

321 Enzyme 1 is introduced to the first tube, cleaving DNA into fragments A and B. AB CD EAD Enzyme 2 is introduced to the next tube, cleaving DNA into fragments C and D. Enzymes 1 & 2 are introduced to the final tube, cleaving DNA into fragments A, E and D.

1 2 1&2 Size Stds. 23,000 bp – 560 bpAgarose Gel A slab of agarose gel has been manufactured with reservoirs to hold the samples of the DNA fragments cleaved by the different restriction enzymes.

Buffer Solution 1 2 1&2 Size Stds. 23,000 bp – 560 bpAgarose Gel The gel is placed in a buffer solution to manage pH balance. pH plays a significant role in electrophoresis.

121&2 Size Stds. 23,000 bp – 560 bp Agarose Gel Buffer Solution When an electrical charge is applied, the negatively charged DNA fragments flow towards the positive charge. DNA with a lesser amount of base pairs flows faster than those of greater base pairs. Base pair sizes of the samples are determined by a control group of size standards (fragments of known measurement) in one of the reservoirs.

Basic Solution 121&2 Size Stds. The basic solution denatures the DNA into single strands Agarose Gel

Salt Solution 121&2 Size Stds. Nylon Filter

Salt Solution 121&2 Size Stds. Paper Towels

Nylon Filter Agarose Gel

Solution with Radioactive Probes

121&2 Size Stds. 23,000 bp – 560 bp Agarose Gel Buffer Solution

Gel Electrophoresis AB CD EAD