CRI DNA sequencing and troubleshooting guide

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Purification before sequencing

  • purify plasmids with QIAprep Spin Miniprep Kit [1]
  • purify PACs with QIAGEN Plasmid Kit [2]
  • purify PCR with products Shrimp Alkaline Phoshatase and Exonuclease I [3]
    • xoSAP-IT NEW 1000 reactions US78200
    • Exonuclease I 2500 reactions E70073Z
    • Shrimp Alkaline Phosphatase 500 reactions E70092Y
  • purify PCR product with QIAGEN QIAquick PCR Purification Kit [4]

Purification after sequencing

On capillary instruments, clean up products with Edge Biosystems: Edge Gel Filtration Cartidge cat# 42453

  1. centrifuge the Edge plates 750 G for 2 min
  2. transfer the cartidge to a clean microcentrifuge tube
  3. add your sample (check fluid runs into the gel)
  4. close cap
  5. centrifuge at 750 G for 2 min