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Finkelstein Lab Wiki

DNA Sequencing

Contents

Section titled “Contents”
  1. Sample requirements
  2. References

1. Sample requirements

Section titled “1. Sample requirements”
  • Plasmid DNA (~5 kb): 15 to 30 ng/µL (180–360 ng total in 12 µL)
  • PCR products: 2 to 5 ng per 100 bp total
  • Primer: 10 pmol of primer (1 µL of 10 µM stock) per sample. Some standard primers are available from the core free of charge.
  • Total volume: Dilute to 12 µL with nuclease-free water in individual 1.5 mL tubes (do not use TE, as EDTA inhibits the sequencing reaction; for Core_Prep_Template-Only samples, use separate tubes for forward and reverse reactions).
  • Labeling: Write the order number and sample number (1 to n) on the top of each tube (do not write the full sample name on the tube).
  • Plates: If submitting 24 samples or more, a 96-well plate is preferred.

2. References

Section titled “2. References”