ABRF '97: Techniques at the Genome/Proteome Interface

Tutorial #6

DNA SEQUENCE ANALYSIS

• Sample Related Problems
  • Template Quantity can Affect Quality of Results
  • Primer Design Problem
  • Sequence Quality Can Be Affected by High Primer Tm
  • Primer Walk with Repeat Sequence
  • Sequencing Through Long PolyT
  • Extra M13 Reverse Priming Site
  • Extra T3 Priming Site
  • Extra T7 Priming Site
  • Miscalled Bases Result in Primer Failure
• Instrument and Software Related Problems
  • Defective Plate
  • Corrupted Gel File
  • Short PCR Products
  • 3.0 Tracking
  • Correct and Incorrect Pixel Positions
  • Peculiar Scan
  • Lane Numbering Glitch
  • Spikes and Noise
  • Warped heat plate
• Gel Related Problems
  • Distorted bands
  • Hourglass gel
  • Haze at the End of the Gel
  • Gel Quality and Position on a Gel Affects Results
  • Red Rain
  • Taurine Buffer Solves Loss of Resolution Between 200 and 300 Bases
• Peak Patterns
  • Peak Patterns Seen Using AmpliTaq® DNA Polymerase, FS
• Comparison of "New" and "Old" Dyes
  • Comparison of dRhodamine dyes with "Old" Rhodamine dyes using TaqFS and Thermo-Sequenase
• Additional trouble shooting tips can be obtained by visiting the University of Leicester DNA sequencing pages.

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