- Sequencing reaction mix
- UH, Manoa Sequencing Service
The ExoSAP reaction "removes" the dNTP's and PCR primers from the PCR product to prepare it for seqeuncing. It contains an exonuclease and shrimp alkaline phosphatase.
- 10 ul PCR product
- 4 ul ExoSAP
The mixture is heated to 37C for 15 minutes then 80C for 15 minutes. It is under the program "EXSAP" on the thermocycler.
The sequencing reaction mix for a 10ul volume is
- 3ul H2O
- 3ul ExoSAP'd PCR product
- 4ul 1uM sequencing primer
We use the University's sequencing service (link), which is currently $2.50 per lane. This form (link) must be filled out and the plate and form dropped off at Snyder 312. We put the reaction for each sample in a small eppendorf tube arranged in order in a plate.
Ordering: (not an endorsement, links to company products are put here for convenience; this list is incomplete and we have not tried and compared many competitive products)
(to be continued...)