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Gel Electrophoresis and Probes (Southern Blotting) Group A,
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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.
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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.
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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.
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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.
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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.
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Basic Solution 121&2 Size Stds. The basic solution denatures the DNA into single strands Agarose Gel
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Salt Solution 121&2 Size Stds. Nylon Filter
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Salt Solution 121&2 Size Stds. Paper Towels
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Nylon Filter Agarose Gel
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Solution with Radioactive Probes
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121&2 Size Stds. 23,000 bp – 560 bp Agarose Gel Buffer Solution
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Gel Electrophoresis AB CD EAD
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