CRI DNA sequencing and troubleshooting guide

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