1 / 19

Gel Electrophoresis of DNA

Gel Electrophoresis of DNA. Start. TABLE OF CONTENTS. Apparatus and Preparation. Results. Apparatus and Preparation. Electrophoresis Apparatus. Loading Gel Beds. Electrophoresis Apparatus. chamber. casting trays. tips. pipet. chamber cover. comb. scoop. Loading Gel Beds. pipet.

ion
Download Presentation

Gel Electrophoresis of DNA

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Gel Electrophoresis of DNA Start

  2. TABLE OF CONTENTS Apparatus and Preparation Results

  3. Apparatus and Preparation Electrophoresis Apparatus Loading Gel Beds

  4. Electrophoresis Apparatus chamber casting trays tips pipet chamber cover comb scoop

  5. Loading Gel Beds pipet loaded well Lane empty well DNA Solute Migration

  6. LOADING THE GEL BED WELL 1 2 DYE MIXTURE SAMPLE LOADED INTO WELL 2 3 4 DNA UNKNOWN SAMPLE LOADED INTO WELL 4 5 6 DNA MARKERS SAMPLE LOADED INTO WELL 7 7 8 continue animation

  7. Gel bed is submerged in a buffer and an electrical current is placed across the chamber. Solutes are pulled through thegel bed toward the pole opposite to their charge. - + WELL 1 Negatively charged DNA fragments and dye mixture solutes will migrate toward the positive pole. 2 3 4 5 6 7 8 continue animation

  8. SOLUTE MIGRATION THROUGH THE GEL BED Dye Mixture Unknown DNA Fragment DNA Markers

  9. Results DNA Solute Banding Measurement of Fragment Migration Interpretation of Data

  10. Gel Bed with DNA Solute Banding Marker Fragment Lane Unknown Fragment Lane continue animation Migration Direction

  11. Immersion of the gel bed in methylene blue allows the DNA fragments to appear as bands Unknown DNA Fragment DNA Markers continue animation

  12. Lighter fragments migrate farther from starting well Heavier fragments do not migrate as far Unknown DNA Fragment Lighter Fragments Heavier Fragments DNA Markers

  13. Measurement of Fragment Migration Distances on Gel Bed Lighter Fragments (Lower Molecular Weight) Heavier Fragments (Higher Molecular Weight) Measure Distances Unknown Fragment continue animation

  14. Measure distance fragments migrate from starting well (in millimeters) ... for Unknown DNA Fragment for DNA Markers continue animation

  15. Fragments migrated (in millimeters) ... 15 for Unknown DNA Fragment 24 18 12 6 5 3 for DNA Markers continue animation

  16. KNOWN MARKERFRAGMENT LENGTH (in kilobase pairs) DISTANCE FRAGMENT MIGRATED ON GEL BED (in millimeters) DNA MARKER FRAGMENT 9.0 3 1 7.0 5 2 6 3 6.5 5.0 12 4 3.0 18 5 2.0 24 6 UNKNOWN DNA FRAGMENT 15 ?

  17. Length of fragment is plotted in kilobase pairs (KBP) along vertical ordinate. Migration of fragment is plotted in millimeters (mm) along horizontal axis. Semilog Graph Paper 2 10 9 Fragment LengthKBP 8 7 6 5 4 3 2 1 28 16 12 20 24 6 10 14 18 22 4 26 8 Fragment Distance Migration (mm) continue animation

  18. Relationship of Fragment Length to Distance Migrated 2 10 9 Fragment LengthKBP Plot data points 8 7 Approximate an area around each point 6 Draw a “best fit” straight line through these areas 5 4 best fit line 3 2 1 28 16 12 20 24 6 5 10 14 18 22 4 26 3 8 Fragment Distance Migration (mm) continue animation

  19. Relationship of Fragment Length to Distance Migrated 2 10 9 Fragment LengthKBP 8 Draw a vertical line up to meet best fit line 7 Draw a horizontal line over to the ordinate 6 5 4 best fit line length of unknownfragment is 3.75 KBP 3 2 Unknown fragment migrated 15 mm 1 28 16 12 20 24 6 5 10 14 18 22 4 26 3 8 Fragment Distance Migration (mm)

More Related