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Biotechniques

Biotechniques. Gel electrophoresis. Separates molecules according to size and charge. Different sized segments of DNA cut by restriction enzymes Segments marked with fluorescent or radioactive dyes Show up as bands on the gel.

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Biotechniques

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  1. Biotechniques

  2. Gel electrophoresis Separates molecules according to size and charge. Different sized segments of DNA cut by restriction enzymes Segments marked with fluorescent or radioactive dyes Show up as bands on the gel.

  3. Precipitation & Centrifugation  DNA fragments form solid pellet at bottom of tube.

  4. Gel electrophoresis machines can sequence up to 600 bases at a time

  5. Loading the Samples • Known controlDNA sample is applied to the first lane • Problems with the control sequence  all results invalid.

  6. Gel electrophoresis -ve +ve Carried out on agarose or acrylamide gel sandwiched between two plates. Smaller fragments moving further in the same time than larger fragments.

  7. Samples placed in wells with buffer Sample Cathode Buffer Electric current Gel Anode Buffer solution

  8. Analyzing DNA Visualizedby applying dyes or radio-labeled probes. DNA markers: DNA molecules with known molecular weights.

  9. Bands visualized under UV light

  10. Gel electrophoresis http://video.google.com/videoplay?docid=-8635131628481451883#docid=5539862268299424434 (electrophoresis, 1 min, good short intro) http://video.google.com/videoplay?docid=-8635131628481451883#docid=8318722871181692802 (Woolfe, electrophoresis, 10 min) http://video.google.com/videoplay?docid=-8635131628481451883# (gel electrophoresis, 7 min – bit slow)

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